(Substituted)acyl dipeptidyl inhibitors of the ICE/CED-3 family of cysteine proteases

ABSTRACT

This invention is directed to novel (substituted) acyl dipeptidyl ICE/ced-3 family inhibitor compounds. The invention is also directed to pharmaceutical compositions containing these compounds, as well as the use of such compositions in the treatment of patients suffering inflammatory, autoimmune and neurodegenerative diseases, for the prevention of ischemic injury, and for the preservation of organs that are to undergo a transplantation procedure.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of pending U.S. Patent ApplicationNo. PCT/US99/24756, filed Oct. 22, 1999; which is a continuation-in-partof U.S. patent application Ser. No. 09/177,546, filed Oct. 22, 1998 andissued as U.S. Pat. No. 6,242,422 on Jun. 5, 2001.

TECHNICAL FIELD

The present invention relates to novel classes of compounds which areinhibitors of interleukin-1β converting enzyme and related proteases(“ICE/ced-3 family of cysteine proteases”), as well as to pharmaceuticalcompositions comprising these compounds and to methods of using suchpharmaceutical compositions.

BACKGROUND OF THE INVENTION

Interleukin 1 (“IL-1”) is a major pro-inflammatory and immunoregulatoryprotein that stimulates fibroblast differentiation and proliferation,the production of prostaglandins, collagenase and phospholipase bysynovial cells and chondrocytes, basophil and eosinophil degranulationand neutrophil activation. Oppenheim, J. H. et al., Immunology Today,7:45–56 (1986). As such, it is involved in the pathogenesis of chronicand acute inflammatory and autoimmune diseases. IL-1 is predominantlyproduced by peripheral blood monocytes as part of the inflammatoryresponse. Mosely, B. S. et al., Proc. Nat. Acad. Sci., 84:4572–4576(1987); Lonnemann, G. et al., Eur. J. Immunol., 19:1531–1536 (1989).

IL-1β is synthesized as a biologically inactive precursor, proIL-1β.ProIL-1β is cleaved by a cysteine protease called interleukin-1βconverting enzyme (“ICE”) between Asp-116 and Ala-117 to produce thebiologically active C-terminal fragment found in human serum andsynovial fluid. Sleath, P. R. et al., J. Biol. Chem., 265:14526–14528(1992); A. D. Howard et al., J. Immunol., 147:2964–2969 (1991).

ICE is a cysteine protease localized primarily in monocytes. In additionto promoting the pro-inflammatory and immunoregulatory properties ofIL-1β, ICE, and particularly its homologues, also appear to be involvedin the regulation of cell death or apoptosis. Yuan, J. et al., Cell,75:641–652 (1993); Miura, M. et al., Cell, 75:653–660 (1993);Nett-Giordalisi, M. A. et al., J. Cell Biochem., 17B:117 (1993). Inparticular, ICE or ICE/ced-3 homologues are thought to be associatedwith the regulation of apoptosis in neurogenerative diseases, such asAlzheimer's and Parkinson's disease. Marx, J. and M. Baringa, Science,259:760–762 (1993); Gagliardini, V. et al., Science, 263:826–828 (1994).

Thus, disease states in which inhibitors of the ICE/ced-3 family ofcysteine proteases may be useful as therapeutic agents include:infectious diseases, such as meningitis and salpingitis; septic shock,respiratory diseases; inflammatory conditions, such as arthritis,cholangitis, colitis, encephalitis, endocerolitis, hepatitis,pancreatitis and reperfusion injury, ischemic diseases such as themyocardial infarction, stroke and ischemic kidney disease; immune-baseddiseases, such as hypersensitivity; auto-immune diseases, such asmultiple sclerosis; bone diseases; and certain neurodegenerativediseases, such as Alzheimer's and Parkinson's disease. Such inhibitorsare also useful for the repopulation of hematopoietic cells followingchemo- and radiation therapy and for prolonging organ viability for usein transplantation.

ICE/ced-3 inhibitors represent a class of compounds useful for thecontrol of the above-listed disease states. Peptide and peptidylinhibitors of ICE have been described. However, such inhibitors havebeen typically characterized by undesirable pharmacologic properties,such as poor oral absorption, poor stability and rapid metabolism.Plattner, J. J. and D. W. Norbeck, in Drug Discovery Technologies, C. R.Clark and W. H. Moos, Eds. (Ellis Horwood, Chichester, England, 1990),pp. 92–126. These undesirable properties have hampered their developmentinto effective drugs.

Accordingly, the need exists for compounds that can effectively inhibitthe action of the ICE/ced-3 family of proteases, for use as agents forpreventing unwanted apoptosis and for treating chronic and acute formsof IL-1 mediated diseases, such as inflammatory, autoimmune orneurodegenerative diseases. The present invention satisfies this needand provides further related advantages.

SUMMARY OF THE INVENTION

In general, the compounds of this invention incorporate an aryl orheteroaryl substituted acyl group as a dipeptide mimetic. The resultingcompounds exhibit improved properties relative to their peptidiccounterparts, for example, such as improved cell penetration or improvedabsorption and metabolic stability resulting in enhancedbioavailability.

One aspect of the instant invention is the compounds of the Formula I:

wherein A, B, X, n, q, R¹, R² and R³ are as defined below, as well aspharmacuetically acceptable salts thereof.

A further aspect of the instant invention is a pharmaceuticalcomposition comprising a compound of the above Formula I and apharmaceutically-acceptable carrier therefor.

Another aspect of this invention involves a method for treating anautoimmune disease comprising administering an effective amount of apharmaceutical composition discussed above to a patient in need of suchtreatment.

Yet another aspect of the instant invention is a method for treating aninflammatory disease comprising administering an effective amount of apharmaceutical composition discussed above to a patient in need of suchtreatment.

A further aspect of the instant invention is a method for treating aneurodegenerative disease comprising administering an effective amountof a pharmaceutical composition discussed above to a patient in need ofsuch treatment.

Another aspect of the instant invention is a method of preventingischemic injury to a patient suffering from a disease associated withischemic injury comprising administering an effective amount of thepharmaceutical composition discussed above to a patient in need of suchtreatment.

A further aspect of the instant invention is a method for expanding ofhematopoietic cell populations and/or enhancing their survival bycontacting the cells with an effective amount of the pharmaceuticalcomposition discussed above. Cell populations included in the method ofthe invention include (but are not limited to) granulocytes, monocytes,erthrocytes, lymphocytes and platelets for use in cell transfusions.

An alternate aspect of the instant invention is a method of prolongingthe viability of an organ that has been removed from the donor for thepurpose of a future transplantation procedure, which comprises applyingan effective amount of the pharmaceutical composition discussed above tothe organ, thereby prolonging the viability of the organ as compared toan untreated organ. The organ may be an intact organ, or isolated cellsderived from an organ (e.g., isolated pancreatic islet cells, isolateddopaminergic neurons, blood or hematopoietic cells).

These and other aspects of this invention will be evident upon referenceto the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, one aspect of the instant invention is the compoundsof the Formula I:

wherein:

n is 0, 1 or 2;

q is 1 or 2;

X is CH₂, C═O, O, S, NH, C═ONH or CH₂OC═ONH;

A is a natural or unnatural amino acid of Formula IIa–i:

B is a hydrogen atom, a deuterium atom, C₁₋₁₀ straight chain or branchedalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, substitutednaphthyl, 2-benzoxazolyl, substituted 2-oxazolyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), (CH₂)_(m)heteroaryl, halomethyl, CO₂R¹³, CONR¹⁴R¹⁵,CH₂ZR¹⁶, CH₂OCO(aryl), CH₂OCO(substituted aryl), CH₂OCO(heteroaryl),CH₂OCO(substituted heteroaryl), or CH₂OPO(R¹⁷)R¹⁸, where Z is an oxygenor a sulfur atom, or B is a group of the Formula IIIa–c:

R¹ is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, or substituted heteroaryl;

R² is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂, (CH₂)_(p)CO₂R₃,(CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or(CH₂)_(m)heteroaryl, wherein heteroaryl includes (but is not limited to)substituted or unsubstituted pyridyl, thienyl, furyl, thiazolyl,imidazolyl, pyrazolyl, isoxazolyl, pyrazinyl, pyrimidyl, triazinyl,tetrazolyl, and indolyl;

R³ is hydrogen, alkyl, cycloalkyl, (cycloalkyl)alkyl, phenylalkyl, orsubstituted phenylalkyl;

and wherein

R⁴ is alkyl, cycloalkyl, phenyl, substituted phenyl, (CH₂)_(m)NH₂,(CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂, (CH₂)_(p)CO₂R³, (CH₂)_(p)OR¹¹,(CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or(CH₂)_(m)heteroaryl, wherein heteroaryl includes (but is not limited to)pyridyl, thienyl, furyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl,pyrazinyl, pyrimidyl, triazinyl, tetrazolyl, and indolyl;

R^(4a) is hydrogen or methyl, or R⁴ and R^(4a) taken together are—(CH₂)_(d)— where d is an interger from 2 to 6;

R⁵ is phenyl, substituted phenyl, (CH₂)_(p)phenyl, (CH₂)_(p)(substitutedphenyl), cycloalkyl, or benzofused cycloalkyl;

R⁶ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl);

R⁷ is hydrogen, fluorine, oxo (i.e., ═O), alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹, SR¹²,or NHCOR¹⁰;

R⁸ is hydrogen, oxo, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R⁹ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or COR¹⁰;

R¹⁰ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹³, or N¹⁴R¹⁵;

R¹¹ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹² is alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl);

R¹³ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹⁴ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, substituted naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹⁵ is hydrogen or alkyl; or

R¹⁴ and R¹⁵ taken together form a five, six or seven memberedcarbocyclic or heterocyclic ring, such as morpholine or N-substitutedpiperazine;

R¹⁶ is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1or 2-naphthyl), or (CH₂)_(m)heteroaryl;

R¹⁷ and R¹⁸ are independently alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, or phenylalkyl, substituted phenylalkyl, or(cycloalkyl)alkyl;

R¹⁹ and R²⁰ are independently hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(m)phenyl, or (CH₂)_(m)(substituted phenyl), or R¹⁹ andR²⁰ taken together are —(CH═CH)₂—;

R²¹ is hydrogen, alkyl, phenyl, substituted phenyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl);

R²², R²³ and R²⁴ are independently hydrogen or alkyl;

Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S;

Y² is O or NR²⁴;

Y³ is CH₂, O, or NR²⁴;

a is 0 or 1 and b is 1 or 2, provided that when a is 1 then b is 1;

c is 1 or 2, provided that when c is 1 then a is 0 and b is 1;

m is 1, 2, 3 or 4; and

p is 1 or 2;

or a pharmaceutically acceptable salt thereof.

As used herein, the term “alkyl” means a straight or branched C₁ to C₃carbon chain such as methyl, ethyl, tert-butyl, iso-propyl, n-octyl, andthe like. The term “lower alkyl” means a straight or branched C₁ to C₆carbon chain, such as methyl, ethyl, iso-propyl, and the like.

The term “cycloalkyl” means a mono-, bi-, or tricyclic ring that iseither fully saturated or partially unsaturated. Examples of such a ringinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,adamantyl, cyclooctyl, cis- or trans decalin, bicyclo[2.2.1]hept-2-ene,cyclohex-1-enyl, cyclopent-1-enyl, 1,4-cyclooctadienyl, and the like.

The term “(cycloalkyl)alkyl” means the above-defined alkyl groupsubstituted with one of the above cycloalkyl rings. Examples of such agroup include (cyclohexyl)methyl, 3-(cyclopropyl)-n-propyl,5-(cyclopentyl)hexyl, 6-(adamantyl)hexyl, and the like.

The term “substituted phenyl” specifies a phenyl group substituted withone or more, and preferably one or two, substituents chosen fromhalogen, hydroxy, protected hydroxy, cyano, nitro, trifluoromethyl,alkyl, alkoxy, acyl, acyloxy, carboxy, protected carboxy, carboxymethyl,protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino,protected amino, (monosubstituted)amino, protected(monosubstituted)amino, (disubstituted)amino, carboxamide, protectedcarboxamide. N-(lower alkyl)carboxamide, protected N-(loweralkyl)carboxamide, N,N-di(lower alkyl)carboxamide, N-((loweralkyl)sulfonyl)amino, N-(phenylsulfonyl)amino or by a substituted orunsubstituted phenyl group, such that in the latter case a biphenyl ornaphthyl group results.

Examples of the term “substituted phenyl” includes a mono- ordi(halo)phenyl group such as 2-, 3- or 4-chlorophenyl,2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 2-, 3- or4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-, 3- or4-fluorophenyl and the like; a mono or di(hydroxy)phenyl group such as2-, 3-, or 4-hydroxyphenyl, 2,4-dihydroxyphenyl, the protected-hydroxyderivatives thereof and the like; a nitrophenyl group such as 2-, 3-, or4-nitrophenyl; a cyanophenyl group, for example, 2-, 3- or4-cyanophenyl; a mono- or di(alkyl)phenyl group such as 2-, 3-, or4-methylphenyl, 2,4-dimethylphenyl, 2-, 3- or 4-(iso-propyl)phenyl, 2-,3-, or 4-ethylphenyl, 2-, 3- or 4-(n-propyl)phenyl and the like; a monoor di(alkoxy)phenyl group, for example, 2,6-dimethoxyphenyl, 2-, 3- or4-(iso-propoxy)phenyl, 2-, 3- or 4-(t-butoxy)phenyl,3-ethoxy-4-methoxyphenyl and the like; 2-, 3- or4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protectedcarboxy)phenyl group such as 2-, 3- or 4-carboxyphenyl or2,4-di(protected carboxy)phenyl; a mono- or di(hydroxymethyl)phenyl or(protected hydroxymethyl)phenyl such as 2-, 3- or 4-(protectedhydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- ordi(aminomethyl)phenyl or (protected aminomethyl)phenyl such as 2-, 3- or4-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono-or di(N-(methylsulfonylamino))phenyl such as 2, 3 or4-(N-(methylsulfonylamino))phenyl. Also, the term “substituted phenyl”represents disubstituted phenyl groups wherein the substituents aredifferent, for example, 3-methyl-4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl,4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,2-hydroxy-4-chlorophenyl, and the like.

The term “phenylalkyl” means one of the above phenyl groups attached toone of the above-described alkyl groups, and the term “substitutedphenylalkyl means that either the phenyl or the alkyl, or both, aresubstituted with one or more of the above-identified substituents.Examples of such groups include 2-phenyl-1-chloroethyl,2-(4′-methoxyphenyl)ethyl, 4-(2′,6′-dihydroxy phenyl)n-hexyl,2-(5′-cyano-3′-methoxyphenyl)n-pentyl, 3-(2′,6′-dimethylphenyl)n-propyl,4-chloro-3-aminobenzyl, 6-(4′-methoxyphenyl)-3-carboxy(n-hexyl),5-(4′-aminomethylphenyl)-3-(aminomethyl)n-pentyl,5-phenyl-3-oxo-n-pent-1-yl, (4-hydroxynapth-2-yl)methyl, and the like.

The term “substituted naphthyl” means a naphthyl group substituted withone or more of the above-identified substituents, and the term “(1 or 2naphyl)alkyl” means a naphthyl attached to one of the above-describedalkyl groups at the 1 or 2 position.

The terms “halo” and “halogen” refer to the fluoro, chloro, bromo oriodo groups. These terms may also be used to describe one or morehalogens, which are the same or different. Preferred halogens in thecontext of this invention are chloro and fluoro.

The term “aryl” refers to aromatic five and six membered carbocyclicrings. Six membered rings are preferred.

The term “heteroaryl” denotes optionally substituted aromaticfive-membered or six-membered heterocyclic rings that have 1 to 4heteroatoms, such as oxygen, sulfur and/or nitrogen atoms, in particularnitrogen, either alone or in conjunction with sulfur or oxygen ringatoms.

The following ring systems are representative examples of theheterocyclic radicals denoted by the term “heteroaryl” (whethersubstitued or unsubstituted): thienyl, furyl, pyrrolyl, pyrrolidinyl,imidazolyl, isoxazolyl. triazolyl, thiadiazolyl, oxadiazolyl,tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl,pyridazinyl, oxazinyl, triazinyl, thiadiazinyl tetrazolo1,5-[b]pyridazinyl and purinyl, as well as benzo-fused derivatives, forexample, benzoxazolyl, benzothiazolyl, benzimidazolyl and indolyl.

Substituents for the above optionally substituted heteroaryl rings arefrom one to three halo, trihalomethyl, amino, protected amino, aminosalts, mono-substituted amino, di-substituted amino, carboxy, protectedcarboxy, carboxylate salts, hydroxy, protected hydroxy, salts of ahydroxy group, lower alkoxy, lower alkylthio, lower alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, (cycloalkyl)alkyl,substituted (cycloalkyl)alkyl, phenyl, substituted phenyl, phenylalkyl,and substituted phenylalkyl groups.

Substituents for the heteroaryl group are as defined above, or as setforth below. As used in conjunction with the above substituents forheteroaryl rings, “trihalomethyl” can be trifluoromethyl,trichloromethyl, tribromomethyl or trilodomethyl, “lower alkoxy” means aC₁ to C₄ alkoxy group, similarly, “lower alkylthio” means a C₁ to C₄alkylthio group. The term “substituted lower alkyl” means theabove-defined lower alkyl group substituted from one to three times by ahydroxy, protected hydroxy, amino, protected amino, cyano, halo,trifluoromethyl, mono-substituted amino, di-substituted amino, loweralkoxy, lower alkylthio, carboxy, protected carboxy, or a carboxy,amino, and/or hydroxy salt.

As used in conjunction with the substituents for the heteroaryl rings,the terms “substituted (cycloalkyl)alkyl” and “substituted cycloalkyl”are as defined above substituted with the same groups as listed for a“substituted alkyl” group. The term “(monosubstituted)amino” refers toan amino group with one substituent chosen from the group consisting ofphenyl, substituted phenyl, alkyl substituted alkyl C₁ to C₇ acyl, C₂ toC₇ alkenyl, C₂ to C₇ substituted alkenyl, C₂ to C₇ alkynyl, C₇ to C₁₆alkylaryl, C₇ to C₁₆ substituted alkylaryl and heteroaryl group. The(monosubstituted)amino can additionally have an amino-protecting groupas encompassed by the term “protected (monosubstituted)amino.” The term“(disubstituted)amino” refers to amino groups with two substituentschosen from the group consisting of phenyl, substituted phenyl, alkyl,substituted alkyl, C₁ to C₇ acyl, C₂ to C₇ alkenyl, C₂ to C₇ alkynyl C₇to C₁₆ alkylaryl, C₇ to C₁₆ substituted alkylaryl and heteroaryl. Thetwo substituents can be the same or different. The term“heteroaryl(alkyl)” denotes an alkyl group as defined above, substitutedat any position by a heteroaryl group, as above defined.

Furthermore, the above optionally substituted five-membered orsix-membered heterocyclic rings can optionally be fused to a aromatic5-membered or 6-membered aryl or heteroaryl ring system. For example,the rings can be optionally fused to an aromatic 5-membered or6-membered ring system such as a pyridine or a triazole system, andpreferably to a benzene ring.

The term “pharmaceutically-acceptable salt” encompasses those salts thatform with the carboxylate anions and includes salts formed with theorganic and inorganic cations such as those chosen from the alkali andalkaline earth metals, (for example, lithium, sodium, potassium,magnesium, barium and calcium); and ammonium ion; and the organiccations (for example, dibenzylammonium, benzylammonium,2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium,phenylethylbenzylammonium, dibenzylethylenediammonium and like cations.)Other cations encompassed by the above term include the protonated formof procaine, quinine and N-methylglucosamine, the protonated forms ofbasic amino acids such as glycine, ornithine, histidine, phenylglycine,lysine, and arginine. Furthermore, any zwitterionic form of the instantcompounds formed by a carboxylic acid and an amino group is referred toby this term. A preferred cation for the carboxylate anion is the sodiumcation. Furthermore, the term includes salts that form by standardacid-base reactions with basic groups (such as amino groups) andincludes organic or inorganic acids. Such acids include hydrochloric,sulfuric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric,palmitic, cholic, pamoic, mucic, D-glutamic, D-camphoric, glutaric,phthalic, tartaric, lauric, stearic, salicyclic, methanesulfonic,benzenesulfonic, sorbic, picric, benzoic, cinnamic, and the like acids.

The compounds of Formula I may also exist as solvates and hydrates.Thus, these compounds may crystallize with, for example, waters ofhydration, or one, a number of, or any fraction thereof of molecules ofthe mother liquor solvent. The solvates and hydrates of such compoundsare included within the scope of this invention.

The term “carboxy-protecting group” as used herein refers to one of theester derivatives of the carboxylic acid group commonly employed toblock or protect the carboxylic acid group while reactions are carriedout on other functional groups on the compound. Examples of suchcarboxylic acid protecting groups include t-butyl, 4-nitrobenzyl,4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl,2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl,3,4-methylenedioxybenzyl, benzhydryl, 4,4′-dimethoxytrityl,4,4′,4″-trimethoxytrityl, 2-phenylpropyl, trimethylsilyl,t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,β-(trimethylsilyl)ethyl, β-(di(n-butyl)methylsilyl)ethyl.p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl,1-(trimethylsilylmethyl)-propenyl and like moieties. The species ofcarboxy-protecting group employed is not critical so long as thederivatized carboxylic acid is stable to the conditions of subsequentreaction(s) and can be removed at the appropriate point withoutdisrupting the remainder of the molecule. Further examples of thesegroups are found in C. B. Reese and E. Haslam, “Protective Groups inOrganic Chemistry,” J. G. W. McOmie, Ed., Plenum Press, New York, N.Y.,1973, Chapter 5, respectively, and T. W. Greene and P. G. M. Wuts,“Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons,New York, N.Y., 1991, Chapter 5, each of which is incorporated herein byreference. A related term is “protected carboxy,” which refers to acarboxy group substituted with one of the above carboxy-protectinggroups.

The term “hydroxy-protecting group” refers to readily cleavable groupsbonded to hydroxyl groups, such as the tetrahydropyranyl,2-methoxyprop-2-yl, 1-ethoxyeth-1-yl, methoxymethyl,β-methoxyethoxymethyl, methylthiomethyl, t-butyl, t-amyl, trityl,4-methoxytrityl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, benzyl,allyl, trimethylsilyl, (t-butyl)dimethylsilyl,2,2,2-trichloroethoxycarbonyl, and the like.

Further examples of hydroxy-protecting groups are described by C. B.Reese and E. Haslam, “Protective Groups in Organic Chemistry,” J. G. W.McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapters 3 and 4,respectively, and T. W. Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis,” Second Edition, John Wiley and Sons, New York, N.Y.,1991, Chapters 2 and 3. A preferred hydroxy-protecting group is thetert-butyl group. The related term “protected hydroxy” denotes a hydroxygroup bonded to one of the above hydroxy-protecting groups.

The term “amino-protecting group” as used herein refers to substituentsof the amino group commonly employed to block or protect the aminofunctionality while reacting other functional groups of the molecule.The term “protected (monosubstituted)amino” means there is anamino-protecting group on the monosubstituted amino nitrogen atom.

Examples of such amino-protecting groups include the formyl (“For”)group, the trityl group, the phthalimido group, the trichloroacetylgroup, the trifluoroacetyl group, the chloroacetyl, bromoacetyl, andiodoacetyl groups, urethane-type protecting groups, such ast-butoxycarbonyl (“Boc”), 2-(4-biphenylyl)propyl-2-oxycarbonyl (“Bpoc”),2-phenylpropyl-2-oxycarbonyl (“Poc”), 2-(4-xenyl)isopropoxycarbonyl,1,1-diphenylethyl-1-oxycarbonyl, 1,1-diphenylpropyl-1-oxycarbonyl,2-(3,5-dimethoxyphenyl)propyl-2-oxycarbonyl (“Ddz”),2-(p-toluyl)propyl-2-oxycarbonyl, cyclopentanyloxycarbonyl,1-methylcyclopentanyl-oxycarbonyl, cyclohexanyloxy-carbonyl,1-methyl-cyclohexanyloxycarbonyl, 2-methylcyclohexanyl-oxycarbonyl,2-(4-toluylsulfonyl)ethoxycarbonyl, 2-(methylsulfonyl)ethoxycarbonyl,2-(triphenylphosphino)-ethoxycarbonyl, 9-fluorenylmethoxycarbonyl(“Fmoc”), 2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl,1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl,5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyl-oxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl,cyclopropylmethoxycarbonyl, isobornyloxycarbonyl,1-piperidyloxycarbonyl, benzyloxycarbonyl (“Cbz”),4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl,α-2,4,5,-tetramethylbenzyl-oxycarbonyl (“Tmz”),4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl,4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl,4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl,4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxycarbonyl,4-(decyloxy)benzyloxycarbonyl and the like; the benzoylmethylsulfonylgroup, the 2,2,5,7,8-pentamethylchroman-6-sulfonyl group (“PMC”), thedithiasuccinoyl (“Dts”) group, the 2-(nitro)phenyl-sulfenyl group(“Nps”), the diphenylphosphine oxide group, and like amino-protectinggroups. The species of amino-protecting group employed is not criticalso long as the derivatized amino group is stable to the conditions ofthe subsequent reaction(s) and can be removed at the appropriate pointwithout disrupting the remainder of the molecule. Preferredamino-protecting groups are Boc, Cbz and Fmoc. Further examples ofamino-protecting groups embraced by the above term are well known inorganic synthesis and the peptide art and are described by, for example,T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapter7, M. Bodanzsky, “Principles of Peptide Synthesis,” 1st and 2nd revisedEd., Springer-Verlag, New York, N.Y. 1984 and 1993, and J. M. Stewartand J. D. Young. “Solid Phase Peptide Synthesis,” 2nd Ed., PierceChemical Co., Rockford, Ill., 1984, E. Atherton and R. C. Shephard,“Solid Phase Peptide Synthesis—A Practical Approach” IRL Press, Oxford,England (1989), each of which is incorporated herein by reference. Therelated term “protected amino” defines an amino group substituted withan amino-protecting group discussed above.

The terms “natural and unnatural amino acid” refers to both thenaturally occurring amino acids and other non-proteinogenic α-aminoacids commonly utilized by those in the peptide chemistry arts whenpreparing synthetic analogues of naturally occurring peptides, includingD and L forms. The naturally occurring amino acids are glycine, alanine,valine, leucine, isoleucine, serine, methionine, threonine,phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine,aspartic acid, asparagine, glutamic acid, glutamine, γ-carboxyglutamicacid, arginine, ornithine and lysine. Examples of unnatural alpha-aminoacids include hydroxylysine, citrulline, kynurenine,(4-aminophenyl)alanine, 3-(2′-naphthyl)alanine, 3-(1′-naphthyl)alanine,methionine sulfone, (t-butyl)alanine, (t-butyl)glycine,4-hydroxyphenyl-glycine, aminoalanine, phenylglycine, vinylalanine,propargyl-gylcine, 1,2,4-triazolo-3-alanine, thyronine,6-hydroxytryptophan, 5-hydroxytryptophan, 3-hydroxy-kynurenine,3-aminotyrosine, trifluoromethylalanine, 2-thienylalanine,(2-(4-pyridyl)ethyl)cysteine, 3,4-dimethoxy-phenylalanine,3-(2′-thiazolyl)alanine, ibotenic acid,1-amino-1-cyclopentane-carboxylic acid, 1-amino-1-cyclohexanecarboxylicacid, quisqualic acid, 3-(trifluoromethylphenyl)alanine,(cyclohexyl)glycine, thiohistidine, 3-methoxytyrosine, norleucine,norvaline, alloisoleucine, homoarginine, thioproline, dehydro-proline,hydroxyproline, homoproline, indoline-2-carboxylic acid,1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,1,2,3,4-tetrahydroquinoline-2-carboxylic acid, α-amino-n-butyric acid,cyclohexylalanine, 2-amino-3-phenylbutyric acid, phenylalaninesubstituted at the ortho, meta, or para position of the phenyl moietywith one or two of the following groups: a (C₁ to C₄)alkyl, a (C₁ toC₄)alkoxy, a halogen or a nitro group, or substituted once with amethylenedioxy group; β-2- and 3-thienylalanine; β-2- and3-furanylalanine; β-2-, 3- and 4-pyridylalanine; β-(benzothienyl-2- and3-yl)alanine; β-(1- and 2-naphthyl)alanine; O-alkylated derivatives ofserine, threonine or tyrosine; S-alkylated cysteine, S-alkylatedhomocysteine, the O-sulfate, O-phosphate and O-carboxylate esters oftyrosine; 3-(sulfo)tyrosine, 3-(carboxy)tyrosine, 3-(phospho)tyrosine,the 4-methane-sulfonic acid ester of tyrosine, 4-methanephosphonic acidester of tyrosine, 3,5-diiodotyrosine, 3-nitrotyrosine, ε-alkyllysine,and delta-alkyl ornithine. Any of these α-amino acids may be substitutedwith a methyl group at the alpha position, a halogen at any position ofthe aromatic residue on the α-amino side chain, or an appropriateprotective group at the O, N, or S atoms of the side chain residues.Appropriate protective groups are discussed above.

Depending on the choice of solvent and other conditions known to thepractitioner skilled in the art, compounds of this invention may alsotake the ketal or acetal form, which forms are included in the instantinvention. In particular, compounds of Formula I in which R³ is ahydrogen atom (i.e., Formula Ia) may exist in the cyclic ketal or acetalform Formula Ia′ shown below:

In addition, it should be understood that the equilibrium forms of thecompounds of this invention may include tautomeric forms. All such formsof these compounds are expressly included in the present invention.

The compounds of this invention may be modified by appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availabilityincrease solubility to allow administration by injection, altermetabolism and alter rate of exertion. In addition, the compounds may bealtered to pro-drug form such that the desired compound is created inthe body of the patient as the result of the action of metabolic orother biochemical processes on the pro-drug. Some examples of pro-drugforms include ketal, acetal, oxime, and hydrazone forms of compoundswhich contain ketone or aldehyde groups, especially where they occur inthe group donated as “A” in Formula I or the modified aspartic acidresidue attached to the group denoted as “A”.

Compounds of this invention with respect to the groups R¹, R², and X inFormula I, include those wherein:

-   -   R¹ is substituted phenyl (such as 2-substituted phenyl),        naphthyl, or substituted naphthyl;    -   R² is hydrogen, lower alkyl (CH₂)_(p)CO₂R³, (CH₂)_(m)phenyl,        (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or        (CH₂)_(m)tetrazolyl, where p is 1 or 2, m is 1 or 2;    -   R³ is hydrogen or alkyl;    -   X is O or NH;    -   q is 1: and    -   n is 0 or 1.

Other compounds of this invention with respect to the groups R¹, R², andX in Formula I, include those wherein:

-   -   R¹ is substituted phenyl, naphthyl, or substituted naphthyl;    -   R₂ is (CH₂)_(m)tetrazolyl, where m is 1 or 2; and    -   X is C═ONH.

Compounds of this invention with respect to the group “A” in Formula I,include those of Formula IIa wherein:

-   -   R⁴ is lower alkyl, cycloalkyl, phenyl, substituted phenyl,        (CH₂)_(m)NH₂, (CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl,        (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1        or 2-naphthyl);    -   R¹¹ is hydrogen, lower alkyl, cycloalkyl, phenyl, substituted        phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,        (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);    -   R¹² is lower alkyl, cycloalkyl, phenyl, substituted phenyl,        naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,        (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);        and    -   m is 1, 2, 3, 4 and p is 1 or 2.

Compounds of this invention with respect to the group “A” in Formula I,also include those of Formula IIb wherein:

-   -   R⁵ is phenyl, substituted phenyl, (CH₂)_(p)phenyl,        (CH₂)_(p)(substituted phenyl), cycloalkyl, or 2-indanyl; and    -   p is 1 or 2.

Another group of compounds with respect to the group “A” in Formula I,include those of Formula IId wherein:

-   -   R⁷ is hydrogen, fluorine, cycloalkyl, phenyl, substituted        phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,        (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹,        or SR¹²;    -   R¹¹ and R¹² are independently cycloalkyl, phenyl, substituted        phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,        (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);        and    -   m is 1, 2, 3 or 4.

A forth group of compounds with respect to the group “A” in Formula I,include those of Formula IIe wherein:

-   -   R⁸ is hydrogen, oxo, cycloalkyl, phenyl, substituted phenyl, or        naphthyl; and    -   Y is CH₂, (CH₂)₂, (CH₂)₃, or S.

Another group of compounds with respect to the group “A” in Formula I,include those of Formula IIh wherein:

-   -   a is 0 and b is 1 or 2.

Compounds of this invention with respect to the group “B” in Formula I,include those wherein:

-   -   B is hydrogen, 2-benzoxazolyl substituted 2-oxazolyl, CH₂ZR¹⁶,        CH₂OCO(aryl), or CH₂OPO(R¹⁷)R¹⁸ where Z is O or S;    -   R¹⁶ is phenyl, substituted phenyl, naphthyl, substituted        naphthyl, heteroaryl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted        phenyl), (CH₂)_(m)(1 or 2-naphthyl), or (CH₂)_(m)heteroaryl;

R¹⁷ and R¹⁸ are independently alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, phenylalkyl, substituted phenylalkyl and(cycloalkyl)alkyl.

Another group of compounds with respect to the group “B” in Formula I,include those of Formula IIIa–c wherein:

-   -   Y² is O or NR²⁴;    -   Y³ is CH₂, O, or NR²⁴;    -   R¹⁹ and R²⁰ are independently hydrogen, alkyl, phenyl, or R¹⁹        and R²⁰ taken together are —(CH═CH)₂—;

R²¹ is hydrogen, alkyl, phenyl, substituted phenyl, (CH₂)_(m)phenyl, or(CH₂)_(m)(substituted phenyl);

-   -   R²², R²³ and R²⁴ are independently hydrogen or alkyl.

The compounds of Formula I may be synthesized using conventionaltechniques as discussed below. Advantageously, these compounds areconveniently synthesized from readily available starting materials. Tothis end, in the following synthetic schemes, q is 1, and correspondingcompounds wherein q is 2 may be made in the same manner by employing thecorresponding ethylene (—CH₂CH₂—) starting material in place of themethylene (—CH₂—) moiety.

One synthetic route for synthesizing the instant compounds is set forthin the following Scheme 1:

In the above Scheme 1, Formula (V), that is H₂N—C, is a modifiedaspartic acid residue of Formulas Va through Vd:

In the above Scheme 1, “PG” stands for an amino protecting group and “A”stands for a natural or unnatural amino acid of formula IIa through IIi,as discussed above. In Formula Vb through Vd, R^(3′) is a carboxylprotecting group as described in the definition of R¹ in Formula I withthe exception that R^(3′) cannot be a hydrogen atom.

The modified aspartic acids of Formula Va–d can be prepared by methodswell known in the art. See, for example, European Patent Application519,748; PCT Patent Application No. PCT/EP92/02472; PCT PatentApplication No. PCT/US91/06595; PCT Patent Application No.PCT/US91/02339; European Patent Application No. 623,592; World PatentApplication No. WO 93/09135; PCT Patent Application No. PCT/US94/08868;European Patent Application No. 623,606; European Patent Application No.618,223; European Patent Application No. 533,226; European PatentApplication No. 528,487; European Patent Application No. 618,233; PCTPatent Application No. PCT/EP92/02472; World Patent Application No. WO93/09135; PCT Patent Application No. PCT/US93/03589; and PCT PatentApplication No. PCT/US93/00481, all of which are herein incorporated byreference.

The coupling reactions carried out under Step A are performed in thepresence of a standard peptide coupling agent such as the combination ofthe combination of dicyclohexylcarbodiimide(DCC) and1-hydroxy-benzotriazole(HOBt), as well as the BOP(benzotriazolyloxy-tris-(dimethylamino)phosphonium hexafluorophosphate)reagent, pyBOP(benzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluorophosphate),HBTU (O-benzotriazolyly-tetramethylisouronium-hexafluorophosphate), andEEDQ (1-ethyloxycarbonyl-2-ethyloxy-1,2-dihydroquinoline) reagents, thecombination of 1-ethyl(3,3′-dimethyl-1′-aminopropyl)carbodiimide (EDAC)and HOBt, and the like, as discussed in J. Jones, “Amino Acid andPeptide Synthesis,” Steven G. Davis ed., Oxford University Press,Oxford, pp. 25–41 (1992); M. Bodanzky, “Principles of PeptideSynthesis.” Hafner et al. ed., Springer-Verlag, Berlin Heidelberg, pp.9–52 and pp. 202–251 (1984); M. Bodanzky, “Peptide Chemistry, APractical Textbook,” Springer-Verlag, Berlin Heidelberg, pp. 55–73 andpp. 129–180; and Stewart and Young, “Solid Phase Peptide Synthesis,”Pierce Chemical Company, (1984), all of which are herein incorporated byreference. The amino protecting group is then removed and the resultingamine is coupled to the (substituted) carboxylic acid of Formula VII(Step B). Again, this coupling reaction uses the standard peptidecoupling reactions mentioned above.

Alternatively, the (substituted)carboxylic acid of Formula VII can becoupled to an amino ester of Formula IX (Step D). Again, this couplingreaction uses the standard peptide coupling reactions mentioned above.In Formula IX, the group R is a carboxyl protecting group such asmethyl, allyl, benzyl or tert-butyl. After removal of the carboxylprotecting group under standard conditions well known in the art, theresulting carboxylic acid is coupled to amine V using the standardpeptide coupling methods described above (Step E).

In the case where the coupling reaction depicted by either Step A orStep E was carried out with the amino alcohol of Formula Vc, the alcoholmoiety must be oxidized to the corresponding carbonyl compound prior toremoval of the protecting groups. Preferred methods for the oxidationreaction include Swern oxidation (oxalyl chloride-dimethyl sulfoxide,methylene chloride at −78° C. followed by triethylamine); andDess-Martin oxidation (Dess-Martin periodinane, t-butanol, and methylenechloride.) The protecting groups contained in substructures of theFormula Va–d, VII and A are removed by methods well known in the art.These reactions and removal of some or all of the protecting groups areinvolved in Step C in the above Scheme 1.

An alternative synthetic route for synthesizing the instant compounds isset forth in the following Scheme 2:

In the above Scheme 2, “PG” stands for an amino protecting group and “A”stands for a natural or unnatural amino acid of formula IIa through IIi,as discussed above. The group R is a carboxyl protecting group such astrimethylsilyl, methyl, allyl, benzyl or tert-butyl.

The coupling reactions carried out under Step F and Step G are performedin the presence of a standard peptide coupling agent as discussed above.In Step G, the amino protecting group must be removed prior to thecoupling step. In Step H the alpha-carboxy protecting group R of thecompound of Formula XIII is selectively removed and the resultingmono-carboxylic acid treated sequentially with diazomethane andhydrobromic acid to give the alpha-bromoketone of Formula XIV.

In Step I, the bromoketone of Formula XIV is treated with either R¹⁶Z-H,(aryl)-CO₂H, (heteroaryl)-CO₂H, or R¹⁷(R¹⁸)PO₂H in the presence of aninorganic base such as potassium carbonate or potassium fluoride in aninert solvent such as dimethyl formamide to give the correspondingcompound of Formula I in which B is CH₂ZR¹⁶, CH₂OCO(aryl),CH₂OCO(heteroaryl), or CH₂OPO(R¹⁷)R¹⁸, respectively. Compounds ofFormula I in which B is a fragment of Formula III may also be preparedin a similar fashion. The protecting groups contained in substructuresof the Formula VII, XI and A are removed by methods well known in theart. These reactions and removal of some or all of the protecting groupsare involved in Step 1 in the above Scheme 2.

An alternative method for the prepartion of compounds of the instantinvention of Formula I in which R³ and B are both hydrogen (i.e.,Formula Ib) is set forth below in Scheme 3:

In Scheme 3, Fmoc is the amino protecting group9-fluorenylmethoxycarbonyl and the shaded circle labeled “PS” representspolystryene resin.

The coupling of the acid of Formula XV to a primary amine on solidsupport, preferably aminomethyl polystyrene, is carried out usingstandard peptide coupling agents, preferably usingbenzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluorophosphate(pyBOP) in a inert solvent such as dimethylformamide or N-methylpyrrolidone (Step J). After removal of the Fmoc protecting group of XVIby treatment with pyrrolidine-dimethylformamide, the resulting amine iscoupled to Fmoc-amino acid of Formula IVa using standard peptidecoupling conditions as discussed above (Step K).

In Step L the Fmoc protecting group of the compound of Formula XVII isremoved again by treatment with with pyrrolidine-dimethylformamide andthe resulting amine coupled to the (substituted)carboxylic acid ofFormula VII again using standard peptide coupling conditions asdiscussed above. The tert-butyl ester of the compound of Formula XVIIIis removed by treatment with trifluoroacetic acid-methylene chloride inthe presence of a trapping agent such as anisole and the resulting acidcleaved from the solid support by treatment with 37% aqueousformaldehyde/acetic acid/tetrahydrofuran/trifluoroacetic acid,preferably in a ratio of 1/1/5/0.025, to give the aspartyl aldehyde ofFormula Ib (Step M).

Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle (hereinafter collectively referred to as“pharmaceutically-acceptable carriers”). Pharmaceutically acceptablecarriers, adjuvants and vehicles that may be used in the pharmaceuticalcompositions of this invention include, but are not limited to, ionexchange, alumina, aluminum stearate, lecithin, serum proteins, such ashuman serum albumin; buffer substances such as the various phosphates,glycine, sorbic acid, potassium sorbate, partial glyceride mixtures ofsaturated vegetable fatty acids; water, salts or electrolytes, such asprotamine sulfate, disodium hydrogen phosphate, potassium hydrogenphosphate, sodium chloride, and zinc salts; colloidal silica, magnesiumtrisilicate, polyvinyl pyrrolidone, cellulose-based substances,polyethylene glycol, sodium carboxymethylcellulose, polyarylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat, and the like.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or by an implanted reservoir. Oral and parenteraladministration are preferred. The term “parenteral” as used hereinincludes subcutaneous, intracutaneous, intravenous, intramuscular,intra-articular, intrasynovial, intrasternal, intrathecal, intralesionaland intracranial injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carrier which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in capsuleform useful diluents include lactose and dried corn starch. When aqueoussuspensions are administered orally, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweeteningand/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible to topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-applied transdermalpatches are also included in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

The compounds of this invention may be used in combination with eitherconventional anti-inflammatory agents or with matrix metalloproteaseinhibitors, lipoxygenase inhibitors and antagonists of cytokines otherthan IL-1β.

The compounds of this invention can also be administered in combinationwith immunomodulators (e.g., bropirimine, anti-human alpha interferonantibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha,diethyldithiocarbamate, tumor necrosis factor, naltrexons and rEPO) orwith prostaglandins, to prevent or combat IL-1-mediated disease symptomssuch as inflammation.

When the compounds of this invention are administered in combinationtherapies with other agents, they may be administered sequentially orconcurrently to the patient. Alternatively, pharmaceutical compositionsaccording to this invention may be comprised of a combination of acompound of Formula I and another therapeutic or prophylactic agentmentioned above.

The disease states which may be treated or prevented by the instantpharmaceutical compositions include, but are not limited to,inflammatory diseases, autoimmune diseases and neurodegenerativediseases, and for inhibiting unwanted apoptosis involved in ischemicinjury, such as ischemic injury to the heart (e.g., myocardialinfarction), brain (e.g., stroke), and kidney (e.g., ischemic kidneydisease). As a consequence of their ability to inhibit apoptosis, thepresent pharmaceutical compositions are also useful for the repopulationof hematopoietic cells of a patient following chemotherapy. Methods ofadministering an effective amount of the above-described pharmaceuticalcompositions to mammals, also referred to herein as patients, in need ofsuch treatment (that is, those suffering from inflammatory diseases,autoimmune diseases, neurodegenerative diseases and for the repopulationof hematopoietic cells in cancer patients who have undergonechemotherapy) are another aspect of the instant invention. Finally, as afurther consequence of their ability to inhibit apoptosis, the instantpharmaceutical compositions may be used in a method to prolong theviability of organs to be used in transplantations.

Inflammatory disease which may be treated or prevented include, forexample, septic shock, septicemia, and adult respiratory distresssyndrome. Target autoimmune diseases include, for example, rheumatoid,arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, insulin-dependentdiabetes mellitus, autoimmune hemolytic anemia, autoimmune neutropenia,thrombocytopenia, chronic active hepatitis, myasthenia gravis andmultiple sclerosis. Target neurodegenerative diseases include, forexample, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson'sdisease, and primary lateral sclerosis. The pharmaceutical compositionsof this invention may also be used to promote wound healing. Targetdiseases associated with harmful, apoptosis, in other words, thoseassociated with ischemic injury, includes myocardial infarction, stroke,and ischemic kidney disease. The pharmaceutical compositions of thisinvention may also be used to treat infectious diseases, especiallythose involved with viral infections.

The term “effective amount” refers to dosage levels of the order of fromabout 0.05 milligrams to about 140 milligrams per kilogram of bodyweight per day for use in the treatment of the above-indicatedconditions (typically about 2.5 milligrams to about 7 grams per patientper day). For example, inflammation may be effectively treated by theadministration of from about 0.01 to 50 milligrams of the compound perkilogram of body weight per day (about 0.5 milligrams to about 3.5 gramsper patient per day).

The amount of the compounds of Formula I that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration. Forexample, a formulation intended for the oral administration of humansmay contain from 0.5 milligrams to 5 grams of a compound of Formula Icombined with an appropriate and convenient amount of apharmaceutically-acceptable carrier which may vary from about 5 to about95 percent of the total composition. Dosage unit forms will generallycontain between from about 1 milligram to about 500 milligrams of anactive compound of Formula I.

It will be understood, however, that the specific “effective amount” forany particular patient will depend upon a variety of factors includingthe activity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, rate of excretion, drug combination and the severity ofthe particular disease undergoing prevention or therapy.

Although this invention focuses on the use of the compounds disclosedherein for preventing and treating IL-1-mediated diseases, the compoundsof this invention can also be used as inhibitory agents for othercysteine proteases.

The compounds of this invention are also useful as commercial reagentswhich effectively bind to the ICE/ced-3 family of cysteine protease orother cysteine proteases. As commercial reagents, the compounds of thisinvention, and their derivatives, may be used to block proteolysis of atarget peptide or may be derivatized to bind to a stable resin as atethered substrate for affinity chromatography applications. These andother uses which characterize commercial cystine protease inhibitorswill be evident to those of ordinary skill in the art.

In order that this invention be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

In the following Examples, proton NMR spectra were obtained at 300 MHz;chemical shifts are quoted downfield from internal tetramethylsilane.

Preparation 1

Preparation of (3S)-Amino-4-Oxobutanoic Acid tert-Butyl EsterSemicarbazone, p-Toluenesulfonate Salt

Part A: N-(Benzyloxycarbonyl)-L-(N′-Methyl-N′-Methoxy)aspartamideβ-(tert-Butyl) Ester

To a solution of N-(benzyloxycarbonyl)-L-aspartic acid-β-(tert-butyl)ester (14.65 g, 45.3 mmol, Bachem) in CH₂Cl₂ (150 mL) at 0° C. (icebath) under a nitrogen atmosphere was added 1-hydroxybenzotriazolehydrate (7.29 g, 47.6 mmol, Aldrich) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(9.55 g, 49.8 mmol, Sigma). After stirring at 0° C. for 15 min.,N,O-dimethylhydroxylamine hydrochloride (5.10 g, 52.3 mmol, Aldrich) andN-methylmorpholine (5.8 mL, 53 mmol, Aldrich) were added. The mixturewas allowed to warm to room temperature over 3 hours then stirred atroom temperature for 16 hours. The solution was concentrated undervacuum and the residue partitioned between ethyl acetate-5% KHSO₄ (200mL each). The organic phase was washed in turn with 5% KHSO₄, saturatedsodium bicarbonate and saturated sodium chloride solutions; dried overanhydrous sodium sulfate and evaporated to an oil. The oil wascrystallized from hexane to give the title product (16.10 g, 97% yield)as a fluffy white crystalline solid. TLC (ethyl acetate), single spot(UV and PMA): Rf=0.37.

A similar procedure to the one above, starting with 29.3 g ofN-(benzyloxycarbonyl)-L-aspartic acid-β-(tert-butyl)ester (2-fold scaleup) gave 31.18 g (94% yield) of the title product.

Part B: (3S)-(Benzyloxycarbonyl)Amino-4-Oxobutanoic Acid tert-ButylEster Semicarbazone

To a solution ofN-(benzyloxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-β-(tert-butyl)ester (15.50 g, 42.3 mmol) in anhydrous ether (400 mL) at 0° C. (icebath) under a nitrogen atmosphere was added dropwise to a 1.0 M solutionof LiAlH₄ in ether (22.0 mL, 22.0 mmol, Aldrich) at such a rate as tokeep the reaction solution temperature between 0–5° C. (addition time15–20 min). After the addition of the lithium aluminum hydride reagentwas complete, the mixture was stirred at 0–5° C. for 1 hr, then quenchedby the dropwise addition of 0.3 N KHSO₄ solution (100 mL). The resultantmixture was transferred to a separatory funnel adding sufficient 5%KHSO₄ solution (75 mL) to dissolve the solids. The organic phase wasseparated and the combined aqueous washes back-extracted with ether (100mL). The combined ether extracts were washed with saturated NaClsolution, dried over anhydrous sodium sulfate and concentrated in vacuowith minimal heating. TLC (ethyl acetate): streaky spot (UV and PMA)Rf=0.48. TLC (methanol/methylene chloride, 1:9) major spot (UV and PMA):Rf=0.75.

The crude aldehyde was immediately taken up in aqueous ethanol (45 mLwater/105 mL alcohol), placed in an ice bath and treated with sodiumacetate (3.82 g, 46.6 mmol) and semicarbazide hydrochloride (5.20 g,46.6 mmol, Aldrich). The mixture was stirred at 0° C. (ice bath) under anitrogen atmosphere for 3 hrs, allowed to warm to room temperature, andstirred overnight (16 hrs). Most of the ethanol was removed under vacuumand the residue partitioned between ethyl acetate and water (100 mLeach). The organic phase was washed sequentially with 5% KHSO₄,saturated sodium bicarbonate and saturated sodium chloride solutions;dried over anhydrous sodium sulfate and evaporated to dryness. The crudeproduct of this reaction was combined with that of two similarprocedures starting with 15.40 g and 4.625 g ofN-(benzyloxycarbonyl)-L-(N′-methyl -N′-methoxy)aspartamide-β-(tert-butylester) (total: 35.525 g, 97 mmol) and these combined products werepurified by flash chromotagraphy on silica gel eluting withacetone/methylene chloride (3:7) then methanol-acetone-methylenechloride (0.5:3:7) to give pure title product (27.73 g, 78.5%) as acolorless foam. TLC (MeOH-CH₂Cl₂, 1:9): single spot (UV and PMA),Rf=0.51.

Part C: (3S)-Amino-4-Oxobutanoic Acid tert-Butyl Ester Semicarbazone,p-Toluenesulfonate Salt

To a solution of (3S)-(benzyloxycarbonyl)amino-4-oxobutanoic acidtert-butyl ester semicarbazone (13.84 g, 38.0 mmol) in absolute ethanol(250 mL) was added 10% Pd/C (1.50 g, Aldrich) and the resulting mixturestirred under an atmosphere of hydrogen (balloon) until TLC(methanol/methylene chloride, 1:9) indicated complete consumption of thestarting material (60 min). Note: It is important to follow thisreaction closely since the product can be over-reduced. The mixture wasfiltered though Celite and evaporated to an oil. The oil was chased withmethylene chloride (2×75 mL) then with methylene chloride/toluene (1:1,75 mL) to give the crude amine as a white crystalline solid. TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) single spot (UV and PMA) Rf=0.24.Note: In this TLC system, any over-reduced product will show upimmediately below the desired product, Rf=0.18 (PMA only).

The crude amine was taken up in CH₃CN (60 mL) and treated with asolution of p-toluenesulfonic acid monohydrate (7.22 g, 38.0 mmol) inacetonitrile (60 mL). The crystalline precipitate was collected, washedwith acetonitrile and ether, and air-dried to give the title compound(13.95 g, 92% yield) as a white, crystalline solid.

The optical purity of this material was checked by conversion to thecorresponding Mosher amide [1.05 equiv(R)-(−)-α-methoxy-α-(trifluoromethyl)phenylacetyl chloride, 2.1equivalents of i-Pr₂NEt in CH₂Cl₂, room temperature, 30 min]. Thedesired product has a doublet at 7.13 ppm (1H, d, J=2.4 Hz, CH═N) whilethe corresponding signal for its diastereomer is at 7.07 ppm. Theoptical purity of the title compound obtained from the above procedureis typically >95:5.

Preparation 2

Preparation of (3S)-3-(9-Fluorenylmethoxycabonyl)Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

Part A: 4-[2′-(N-t-Butoxycarbonyl)Aminoethyl]Phenoxyacetic Acid, MethylEster

To a suspension 4-hydroxy-phenethylamine (7.00 g, 51.1 mmol, Aldrich) indry dimethylformamide (50 mL) at room temperature under nitrogen wasadded di-tert-butyl dicarbonate (11.0 g, 50.5 mmol). After stirring atroom temperature for 1 hr, the resulting clear solution was treated withmethyl bromoacetate (7.5 mL, 79 mmol) and cesium carbonate (17.5 g, 53.7mmol). After stirring at room temperature for 16 hrs, TLC (Et₂O-toluene;2:8) shows some unalkylated material remained (Rf=0.43) and a secondportion of methyl bromoacetate (2.0 mL, 21 mmol) and cesium carbonate(4.5 g, 14 mmol) were added. After stirring for an additional 24 hrs,the mixture was partitioned between EtOAc-water (250 mL each), organicphase washed succesively with water (3×), 5% potassium bisulfate andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. Trituration of the residue with hexane gave 15.87 g of a tansolid. Filtration of the crude product through a pad of silica geleluting with EtOAc-hexane (2:8) and crystallization from hexane gave thetitle compound (14.75, 93%) as a white granular, crystalline solid. TLC(Et₂O-toluene; 2:8) Rf=0.53.

Part B: 4-(2′-Aminoethyl)Phenoxyacetic Acid, Methyl Ester, Hydrochloride

To a solution 4-[2′-(N-t-butoxycarbonyl)aminoethyl]phenoxyacetic acid,methyl ester (18.31 g, 59.3 mmol) in dioxane (55 mL) at room temperaturewas added 4.0 N HCl in dioxane (55 mL). After stirring at roomtemperature for 16 hrs, the mixture was diluted with Et₂O, theprecipatate collected, washed throughly with Et₂O and dried in vacuo togive the title compound (14.55 g, 94%) was a fluffy white, crystallinesolid.

Part C:1-tert-Butoxycarbonyl-Semicarbazidyl-4-[2′-(4-Ethyl-PhenoxyaceticAcid)]Methyl Ester

A solution of t-butyl carbazate (6.60 g, 50 mmol) in dimethylformamide(50 mL) was added dropwise to a solution carbonyldiimidazole (8.10 g, 50mmol) in dimethylformamide (80 mL) over 40 min at room temperature undernitrogen. After stirring at room temperature for an additional 30 min,4-(2′-aminoethyl)phenoxyacetic acid, methyl ester, hydrochloride (12.3g, 50 mmol) was added as a solid in one portion followed by atriethylamine (8.0 mL, 58 mmol) added dropwise over 30 min. Afterstirring at room temperature for 18 hrs, the mixture was partitionedbetween EtOAc-water (300 mL each). The organic phase was washedsuccesively with water (3×), 5% potassium bisulfate, saturated sodiumbicarbonate, and saturated NaCl solutions, dried over anhydrous Na₂SO₄and evaporated to dryness. Crystallization of the residue fromEtOAc-hexane gave the title compound (15.50, 84%) as an off-whitecrystalline solid. TLC (MeOH-CH₂Cl₂; 1:9) Rf=0.45.

Part D:1-tert-Butoxycarbonyl-Semicarbazidyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

A solution of1-tert-butoxycarbonyl-semicarbazidyl-4-[2′-(4-ethyl-phenoxyaceticacid)]methyl ester (14.68 g, 40 mmol) in dioxane (50 mL) at roomtemperature under nitrogen was added 1.0 N LiOH solution (50 mL). Afterstirring at room temperature for 1 hr, the mixture was acidified withconc. HCl and extracted with EtOAc (100 mL). The organic phase waswashed with saturated NaCl solution, dried over anhydrous Na₂SO₄ andevaporated to a white solid. Recrystallization of the crude product fromTHF-EtOAc-hexane gave the title compound (13.44, 95%) as a whitecrystalline solid. TLC (AcOH-MeOH-CH₂Cl₂; 1:1:8) Rf=0.31.

Part E: Semicarbazidyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]Hydrochloride

To a solution of1-tert-butoxycarbonyl-semicarbazidyl-4-[2′-(4-ethyl-phenoxyacetic acid)](13.43 g, 38.0 mmol) in dioxane (80 mL)-anisole (15 mL) at roomtemperature was added 4.0 N HCl in dioxane (35 mL). After stirring atroom temperature for 18 hrs, additional 4.0 N HCl in dioxane (15 mL) wasadded. After an additional 6 hrs, the precipatate was collected, washedthroughly with dioxane then Et₂O and dried in vacuo to give the titlecompound (11.67 g, 100%) was a white, crystalline solid.

Part F:N-(9-Fluorenylmethoxycarbonyl)-L-(N′-Methyl-N′-Methoxy)aspartamideβ-(tert-Butyl)Ester

To a solution of N-(9-fluorenylmethoxycarbonyl)-L-asparticacid-β-(tert-butyl)ester (16.48 g, 40 mmol) in CH₂Cl₂ (80mL)-tetrahydrofuran (20 mL) at 0° C. (ice bath) under a nitrogenatmosphere was added 1-hydroxybenzotriazole hydrate (7.12 g, 46.5 mmol)followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (9.20 g, 48 mmol). After stirring at 0° C. for 15 min.,N,O-dimethylhydroxylamine hydrochloride (4.68 g, 48 mmol) andN-methylmorpholine (5.2 mL, 47 mmol) were added. The mixture was allowedto warm to room temperature over 2 hours then stirred at roomtemperature for 16 hours. The solution was concentrated under vacuum andthe residue partitioned between ethyl acetate-5% KHSO₄ (200 mL each).The organic phase was washed succesively with 5% KHSO₄, saturated sodiumbicarbonate and saturated sodium chloride solutions; dried overanhydrous sodium sulfate and evaporated to an oil. Purification of thecrude product by flash chromatography on silica gel eluting withEtOAc-hexane (30:70 then 35:65) gave the title product (17.75 g, 98%yield) as a colorless foam. TLC (EtOAc-hexane; 1:1) Rf=0.35.

Part G: (3S)-3-(9-Fluorenylmethoxycabonyl)Amino-4-Oxobutanoic Acidtert-Butyl Ester Semicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

To a solution ofN-(9-fluorenylmethoxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-β-(tert-butyl)ester(13.20 g, 29 mmol) in anhydrous ether (250 mL) at 0° C. (ice bath) undera nitrogen atmosphere was added dropwise to a 1.0 M solution of LiAlH₄in ether (14.5 mL, 14.5 mmol) at such a rate as to keep the reactionsolution temperature between 0–5° C. (addition time 15–20 min). Afterthe addition of the lithium aluminum hydride reagent was complete, themixture was stirred at 0–5° C. for 1 hr, then quenched by the dropwiseaddition of 0.3 N KHSO₄ solution (100 mL). After adding sufficient 0.3 NKHSO₄ solution to dissolve most of the inorganic salts, the mixture wastransferred to a seperatory funnel. The organic phase was separated andthe aqueous phase back-extracted with ether (100 mL). The combined etherextracts were washed with saturated NaCl solution, dried over anhydroussodium sulfate and concentrated in vacuo with minimal heating. TLC(EtOAc-hexane): Rf=0.40.

The crude aldehyde was immediately taken up in ethanol (105 mL)-water(45mL)-tetrahydrofuran(75 mL), placed in an ice bath and treated withsodium acetate (3.20 g, 39 mmol) andsemicarbazidyl-4-[2′-(4-ethyl-phenoxyacetic acid)]hydrochloride (8.65 g,30 mmol). The mixture was stirred at 0° C. (ice bath) under a nitrogenatmosphere for 3 hrs, allowed to warm to room temperature, and stirredovernight (16 hrs). The mixture was concentrated on a rotovap, dilutedwith water and resulting precipitate collected by suction. The materialwas dried in vacuo to give 18.36 g of crude product as a white solid.The crude product of this reaction was combined with that of a smallerscale reaction (6.34 g) starting with 4.55 g (10 mmol) ofN-(9-fluorenylmethoxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-β-(tert-butylester) and partitioned between ethyl acetate-tetrahydrofuran(1:1) and 5%KHSO₄. The organic phase was washed with 5% KHSO₄ and saturated sodiumchloride solutions, dried over anhydrous sodium sulfate and evaporatedto dryness. The residue was purified by filtration through a pad ofsilica gel eluting with terahydrofuran/methylene chloride (1:1). Thecombined product-containing fractions were evaporated to dryness andrecrystallized from tetrahydrofuran-Et₂O to give pure title product(17.01 g, 69%) as a white solid. TLC (AcOH-MeOH-CH₂Cl₂, 1:1:40):Rf=0.19.

Preparation 3 Assay for Inhibition of ICE/ced-3 Protease Family Activity

A. Determination of IC₅₀ Values

Fluorescence enzyme assays detecting the activity of the compounds ofFormula 1 utilizing the recombinant ICE and CPP32 enzymes are performedessentially according to Thornberry et al. (Nature, 356:768:774 (1992))and Nicholson et al. (Nature, 376:37–43 (1995)) respectively, (hereinincorporated by reference) in 96 well microtiter plates. The substrateis Acetyl-Tyr-Val-Ala-Asp-amino-4-methylcoumarin (AMC) for the ICE assayand Acetyl-Asp-Glu-Val-Asp-amino-4-methylcoumarin for the CPP32, Mch2,Mch3 and Mch5 assays. Enzyme reactions are run in ICE buffer (25 mMHEPES, 1 mM EDTA, 0.1% CHAPS, 10% sucrose, pH 7.5) containing 2 mM DTTat room temperature in duplicate. The assays are performed by mixing thefollowing components:

-   -   50 μL ICE, Mch2, Mch5, CPP32 (18.8, 38, 8.1 and 0.153 nM        concentrations, respectively) or Mch3 (1 unit) enzyme in ICE        buffer containing either 8.0 (ICE, Mch2, Mch3, CPP32) or 20        (Mch5) mM DTT;    -   50 μL compound of Formula 1 or ICE buffer (control); and    -   100 μL of 20 μM substrate.

The enzyme and the compound of Formula I to be assayed are allowed topreincubate in the microtitre plate wells for 30 minutes at roomtemperature prior to the addition of substrate to initiate the reaction.Fluorescent AMC product formation is monitored for one hour at roomtemperature by measuring the fluorescence emission at 460 nm using anexcitation wavelength of 360 nm. The fluorescence change in duplicate(control) wells are averaged and the mean values are plotted as afunction of inhibitor concentration to determine the inhibitorconcentration producing 50% inhibition (IC₅₀). The results of this assayare set forth below in Table 1 and in Table 3 (for Table 3, see Examples11 through 52).

The reference compound for this assay was Cbz-ValAlaAsp-H and the valuesare denoted in Table 1 as “Reference”.

TABLE 1 mICE CPP32 MCH-2 MCH-3 MCH-5 Ex. No. IC₅₀(μM) IC₅₀(μM) IC₅₀(μM)IC₅₀(μM) IC₅₀(μM) 1 0.535 0.141 0.995 1.56 0.680 2 0.336 0.355 >10 2.101.20 3 2.55 0.021 0.015 0.587 0.012 4 4.86 0.0038 0.0035 0.130 0.031 52.96 0.401 3.61 10.9 0.733 6 0.385 0.054 1.43 1.65 0.048 7 1.89 0.7311.90 17.0 0.200 8 0.033 0.013 0.037 1.32 0.0076 9 0.087 0.512 0.310 7.240.017 10 6.34 0.241 13.1 2.32 6.34 179 0.204 14.0 3.53 >50 1.55 1860.298 25.3 >50 >50 39.8 188 0.127 0.207 1.01 11.0 0.615 reference 0.06447.0 >10 >10 2.96B. Determination of the Dissociation Constant Ki and Irreversible RateConstant k₃ for Irreversible Inhibitors

For the irreversible inhibition of a ICE/ced-3 Family Protease enzymewith a competitive irreversible inhibitor; using the model representedby the following formulas:

The product formation at time t may be expressed as:

$\begin{matrix}{\lbrack P\rbrack_{t} = {\lbrack E\rbrack^{T}( \frac{\lbrack S\rbrack K_{i}}{\lbrack I\rbrack K_{s}} ){( \frac{k_{s}}{k_{3}} )\lbrack {1 - {\mathbb{e}}^{{- k_{3}}{t/{({1 + {\frac{K_{i}}{\lbrack I\rbrack}{({1 + \frac{\lbrack S\rbrack}{K_{s}}})}}})}}}} \rbrack}}} & {{Equation}\mspace{20mu} 1}\end{matrix}$where E, I, EI and E-I denote the active enzyme, inhibitor, non-covalentenzyme-inhibitor complex and covalent enzyme-inhibitor adduct,respectively. The K_(i) value is the overall dissociation constant ofthe reversible binding steps, and k₃ is the irreversible rate constant.The [S] and K_(s) values are the substate concentration and dissociationconstant of the substrate bound to the enzyme, respectively. [E]^(T) isthe total enzyme concentration.

The above equations were used to determine the K_(i) and k₃ values of agiven inhibitor bound to a ICE/ced-3 family protease. Thus, a continuousassay was run for sixty minutes at various concentrations of theinhibitor and the substrate. The assay was formulated essentially thesame as described above for generating the data in Table 1, except thatthe reaction was initiated by adding the enzyme to thesubstrate-inhibitor mixture. The K_(i) and k₃ values were obtained bysimulating the product AMC formation as a function of time according toEquation 1. The results of this second assay are set forth below inTable 2.

The reference compound for this assay was Cbz-ValAlaAsp-CH₂F and thevalues are denoted in Table 2 as “Reference”. The K_(i) values in Table2 are in micromolar (μM). The k₃/K_(i) values are in moles⁻¹ seconds⁻¹(M⁻¹s⁻¹).

TABLE 2 mICE CPP32 MCH-2 MCH-5 Ex. No. Ki k₃/Ki Ki k₃/Ki Ki k₃/Ki Kik₃/Ki 53 0.053 129,000 0.079 207,000 0.038 36,800 0.040 71,700 54 1.098,280 0.209 59,300 0.057 64,400 0.059 32,300 55 0.246 33,200 0.18641,300 0.039 59,400 0.056 20,400 56 0.324 15,400 0.138 105,000 0.05350,000 0.085 12,800 57 0.120 37,400 0.042 177,000 0.030 91,000 0.06615,900 59 0.184 46,300 0.942 14,400 0.071 10,100 0.090 17,600 60 0.37333,500 0.758 8,440 ND ND 0.467 5,780 61 0.148 93,200 0.360 28,300 ND ND0.217 10,100 62 0.253 45,400 0.052 169,000 0.042 44,000 0.048 18,200 630.079 52,100 0.012 725,000 0.012 56,900 0.012 17,000 64 0.262 3,6300.062 19,200 0.153 2,400 0.235 4,260 65 0.305 6,020 0.102 26,800 0.336 00.354 230 66 0.442 2,700 0.121 17,800 0.344 48 0.406 460 67 0.218 9,1200.033 8,560 0.203 0 0.255 700 68 0.355 14,800 0.110 28,800 0.383 1,6100.821 200 69 0.615 8,400 0.092 21,700 0.951 0 1.30 630 70 0.399 12,1000.104 49,000 0.357 1,330 0.760 480 71 0.193 53,900 0.039 200,000 0.0389,980 0.120 9,100 72 0.718 1,620 0.090 6,460 1.16 90 1.04 120 73 0.5922,170 0.106 9,240 0.862 110 1.03 150 74 0.280 11,900 0.135 35,800 1.25250 1.08 770 75 0.147 14,700 0.061 60,100 0.221 1,510 0.794 1,470 760.090 47,100 0.063 188,000 0.058 81,700 0.081 17,000 77 0.262 11,5000.123 24,400 0.526 630 1.50 670 78 0.137 20,700 0.038 114,000 0.0815,140 0.202 9,080 79 0.091 77,500 0.042 268,000 0.006 78,900 0.03430,200 80 0.926 4,700 0.099 56,600 0.023 13,600 0.146 8,040 103 0.063143,000 0.038 351,000 0.038 39,700 0.025 59,300 104 0.133 50,600 0.054151,000 0.037 50,200 0.059 15,500 105 0.413 18,000 0.341 44,900 0.2336,090 0.160 3,700 106 0.167 42,500 0.048 155,000 0.080 52,900 0.13410,500 107 0.066 106,000 0.014 424,000 0.021 187,000 0.048 27,800 1080.147 37,900 0.041 140,000 0.037 60,400 0.105 9,890 109 0.453 15,5000.136 48,300 0.119 16,800 0.219 4,070 110 0.059 64,900 0.035 272,0000.015 150,000 0.043 18,800 111 0.308 6,500 0.220 21,900 2.16 230 2.87170 115 0.324 8,740 0.046 127,000 0.054 0 4.67 0 121 0.242 24,800 0.047114,000 0.120 5,150 0.276 3,200 128 0.213 5,480 0.254 5,240 2.41 83 4.480 143 0.205 28,300 0.050 121,000 0.028 8,500 0.037 14,500 144 0.12642,500 0.054 144,000 0.070 5,800 0.155 6,340 150 0.263 43,700 0.016698,000 0.009 400,000 0.127 9,340 151 0.349 29,600 0.032 257,000 0.02388,100 0.270 5,900 152 0.191 29,300 0.029 241,000 0.011 191,000 0.06616,600 155 0.168 59,800 0.047 206,000 0.015 166,000 0.136 7,910 1560.438 20,200 0.148 49,700 0.052 14,900 0.293 3,990 157 0.225 39,3000.257 53,300 0.022 72,000 0.072 11,600 158 0.168 34,300 0.109 98,2000.022 103,000 0.264 1,610 159 1.37 4,580 1.18 11,700 0.113 15,000 10.786 160 1.18 11,400 0.132 33,000 0.093 36,600 0.351 3,680 161 0.09886,400 0.019 319,000 0.030 149,000 0.105 15,200 162 0.319 22,200 0.044246,000 0.029 104,000 0.128 5,290 163 0.415 37,800 0.023 308,000 0.012110,000 0.252 7,960 164 0.467 24,000 0.063 137,000 0.023 91,700 0.2236,190 165 0.396 25,500 0.020 335,000 0.008 116,000 0.089 13,100 1660.042 149,000 0.028 317,000 0.011 146,000 0.028 80,000 167 0.501 21,3000.089 56,200 0.042 52,500 0.126 13,600 174 0.779 6,320 1.15 8,210 0.2227,720 1.19 1,260 175 2.34 4,000 1.10 10,900 0.149 20,600 0.377 4,090 1760.480 11,100 3.08 4,330 1.26 1,330 1.16 684 177 0.225 45,600 0.08689,700 0.047 21,100 0.439 4,370 refer. 0.015 214,000 0.820 12,200 0.5942,950 0.018 83,300

The following are examples of compounds of the invention.

EXAMPLE 1

(3S)-3-[N-((1-Naphthyloxy)Acetyl)Leucinyl]Amino-4-Oxobutanoic Acid

Part A: (3S)-3-[(N-Benzyloxycarbonyl)Leucinyl]Amino-4-Oxobutanoic Acidtert-Butyl Ester Semicarbazone

To a solution of (N-benzyloxycarbonyl)leucine N-hydroxysuccinimde ester(1.81 g, 5.0 mmol) in CH₂Cl₂ (30 mL) at room temperature under nitrogenwas added (3S)-amino-4-oxobutanoic acid tert-butyl ester semicarbazone,p-toluenesulfonate salt (2.58 g, 6.4 mmol) followed by diisopropylethylamine (1.2 mL, 6.9 mmol). After stirring at room temperature for 16hrs, the mixture was concentrated and the residue partitioned betweenEtOAc-5% KHSO₄. The organic phase was washed with 5% KHSO₄, saturatedNaHCO₃ and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to give the title compound (2.798 g) as a pale yellow foam.TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.52.

Part B: (3S)-3-(Leucinyl)Amino-4-Oxobutanoic Acid tert-Butyl EsterSemicarbazone

To a solution of crude(3S)-[(N-benzyloxycarbonyl)leucinyl]amino-4-oxobutanoic acid tert-butylester semicarbazone (2.798 g, ca 5.0 mmol) in absolute EtOH (40 mL) wasadded 10% Pd—C (0.40 g) and resulting mixture stirred under a hydrogenatmosphere (balloon) for 1.5 hrs. The mixture was filtered throughCelite washing the filter cake with CH₂Cl₂ and the combined filtratesevaporated to dryness. The residue was chased with CH₂Cl₂ (2×20 mL) togive the title product (2.113 g) as a colorless foam. TLC(MeOH-CH₂Cl₂;1:9) Rf=0.23.

Part C: (3S)-3-[N-((1-Naphthyloxy)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of (1-naphthyloxy)acetic acid (0.150 g, 0.74 mmol) and(3S)-3-(leucinyl)amino-4-oxobutanoic acid tert-butyl ester semicarbazone(0.360 g, ca 0.83 mmol) in N-methylpyrrolidone(2.0 mL)-CH₂Cl₂(2.0 mL) at0° C. (ice bath) under nitrogen was added hydroxybenzotriazole hydrate(0.130 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.195 g, 1.02 mmol). After stirring at 0° C. for 1 hrs and at roomtemperature for 5 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The crude product was purified by flash chromatography elutingwith MeOH-CH₂Cl₂ (2:100 then 5:100) to give the title compound (0.366 g,94%) as a colorless foam. TLC(MeOH-CH₂Cl₂; 5:95) Rf=0.20.

Part D: (3S)-3-[N-((1-Naphthyloxy)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution of(3S)-3-[N-((1-naphthyloxy)acetyl)leucinyl]amino-4-oxobutanoic acidtert-butyl ester semicarbazone (0.366 g 0.69 mmol) in CH₂Cl₂(2.0mL)-anisole(0.5 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (2.0 mL). The resulting solution was stirred atroom temperature for 3 hrs, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (0.354 g, 100%) as an off-white solid.TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.25. TLC(EtOAc-pyridine-AcOH-H₂O;60:20:5:10) Rf=0.48.

Part E: (3S)-3-[N-((1-Naphthyloxy)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid

A solution of(3S)-3-[N-((1-naphthyloxy)acetyl)leucinyl]amino-4-oxobutanoic acidsemicarbazone (0.320 g, 0.68 mmol) in 37% aqueous formaldehyde(1.0mL)-acetic acid(1.0 mL)-methanol(3.0 mL) was stirred at room temperatureunder nitrogen for 3.5 hrs. The resulting solution was diluted withwater and extracted with EtOAc. The extract was washed with water andsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was taken up in EtOAc, filtered through Celite andevaporated to dryness. The product was taken up in a small amount ofdioxane, diluted with water, frozen and lyophilized to give the titlecompound (0.222 g, 79%) as a fluffy white solid.TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.65.

EXAMPLE 2

(3S)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-4-Oxobutanoic Acid

Part A: (3S)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-4-Oxobutanoic Acidtert-Butyl Ester Semicarbazone

To a solution of (N-benzyloxycarbonyl)valine (2.035 g, 8.10 mmol) inCH₂Cl₂(80 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (1.15 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(2.33 g, 12.2 mmol). After stirring at 0° C. for 10 min,(3S)-amino-4-oxobutanoic acid tert-butyl ester semicarbazone,p-toluenesulfonate salt (3.26 g, 8.10 mmol) followed byN-methylmorpholine (0.89 mL, 8.10 mmol) was added. After stirring at 0°C. for 2 hrs and at room temperature for 20 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to dryness. The crude product waspurified by flash chromatography eluting with MeOH-CH₂Cl₂ (2:100 then5:100) to give the title compound (3.50 g, 93%) as a colorless foam.TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.59.

Part B: (3S)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Starting with (3S)-3-[(N-benzyloxycarbonyl)valinyl]-amino-4-oxobutanoicacid tert-butyl ester semicarbazone and follwing the general methoddescribed in Example 1, Parts B through E, the title compound was alsoprepared. TLC(AcOH-MeOH-CH₂Cl₂; 1:20) Rf=0.20. MS(ES) for C₂₁H₂₄N₂O₆ (MW400.61): positive 401(M+H); negative 399(M−H).

EXAMPLE 3

(3S,2′S)-3-[N-((2′-(1-Naphthyloxy)-4′-Carboxy)Butyryl)Leucinyl]Amino-4-OxobutanoicAcid

Part A: (2R)-2-Bromo-4-Carbobenzyloxy-Butyric Acid Methyl Ester

To a solution of D-glutamic acid γ-benzyl ester (5.00 g, 21 mmol) andKBr (7.5 g, 63 mmol) in 2.5N H₂SO₄ (35 mL) at 0° C. (ice bath) was addedNaNO₂ (2.45 g, 35.5 mmol) in small portions over 1.5 hrs keeping theinternal temperature below 5° C. The resulting mixture was stirred at 0°C. for 1 hr and at room temperature for 45 min. The mixture wasextracted with Et₂O, extract washed with water and saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was taken up in CH₂Cl₂, re-dried over Na₂SO₄ and evaporated todryness to give crude (2R)-2-bromo-4-carbobenzyloxy-butyric acid (5.52g) as a colorless oil.

A portion of the crude acid (3.05 g) was taken up in Et₂O (35 mL) andtreated with excess diazomethane (prepared from 3.2 g of1-methyl-3-nitro-1-nitrosoguanidine, 10 mL 40% KOH/35 ml Et₂O at 0° C.)in portions at 0° C. (ice bath). When TLC indicated consumption of theacid material the excess diazomethane was discharged with a few drops ofacetic acid and the mixture was evaporated to a colorless oil. The crudeproduct was purified by flash chromatography on silica gel eluting withEt₂O-hexane(1:9) to give the title compound (1.87 g, 51% overall) as acolorless liquid. TLC(Et₂O-hexane; 35:65) Rf=0.50.

Part B: (2S)-2-(1-Naphthyloxy)-4-Carbobenzyloxy-Butyric Acid MethylEster

To a solution of (2R)-2-bromo-4-carbobenzyloxy-butyric acid methyl ester(1.00 g, 3.17 mmol) and 1-naphthol (0.500 g, 3.47 mmol) indimethylformamide (4.0 mL) at room temperature under nitrogen was addedpowdered anhydrous K₂CO₃ (0.560 g, 4.7 mmol). After stirring at roomtemperature for 3.5 hrs, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, 5% KHSO₄ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa yellow oil. The crude product was purified by flash chromatography onsilica gel eluting with Et₂O-hexane(1:4) to give the title compound(1.105 g, 92%) as a pale yellow oil. TLC(Et₂O-hexane; 3:7; 2developments) Rf=0.33 (bromide Rf=0.52; 1-naphthol Rf=0.41).

Part C: (2S)-2-(1-Naphthyloxy)-4-Carbo(tert-Butoxy)-Butyric Acid MethylEster

A solution of (2S)-2-(1-naphthyloxy)-4-carbobenzyloxy-butyric acidmethyl ester (1.09 g, 2.88 mmol) in MeOH (10 mL) was treated with 10%Pd—C (0.13 g) and stirred under a hydrogen atmosphere (balloon) for 1.5hrs. The mixture was filtered through Celite washing the filter cakewith CH₂Cl₂ and the combined filtrates evaporated to dryness. Theresidue was chased with toluene (10 mL) and CH₂Cl₂ (2×20 mL) to give themonoacid (0.889 g) as a colorless, viscous oil. TLC(MeOH-CH₂Cl₂; 5:95)Rf=0.25.

A solution of the crude acid (0.889 g, ca 2.88 mmol) in drytetrahydrofuran (18 mL) at room temperature under nitrogen was treatedwith triethylamine (0.64 mL, 4.6 mmol) and 2,4,6-trichlorobenzoylchloride (0.605 mL, 3.87 mmol). After stirring at room temperature for18 hrs, the mixture was diluted with Et₂O, filtered through sincteredglass and evaporated to dryness. The crude mixed anhydride was taken upin CH₂Cl₂ (12 mL) and tert-butanol (3.5 mL), and treated with4-dimethylaminopyridine (0.445 g, 3.65 mmol). After stirring at roomtemperature under nitrogen for 3.5 hrs, the mixture was concentrated andpartitioned between EtOAc-5% KHSO₄. The organic phase was washed with 5%KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to an oil. The crude product waspurified by flash chromatography eluting with Et₂O-hexane (1:9) to givethe title compound (0.817 g, 82% overall) as a colorless, viscous oil.TLC(Et₂O-hexane; 3:7) Rf=0.30.

Part D:(3S,2′S)-3-[N-((2′-(1-Naphthyloxy)-4′-Carboxy)Butyryl)Leucinyl]Amino-4-OxobutanoicAcid Semicarbazone Di-tert-Butyl Ester

To a solution of (2S)-2-(1-naphthyloxy)-4-carbo(tert-butoxy)-butyricacid methyl ester (0.136 g, 0.395 mmol) in dioxane(1.5 mL)-water(0.5 mL)at 0° C. (ice bath) under nitrogen, was added 1.0N LiOH solution (0.52mL, 0.52 mmol). After stirring at 0° C. for 30 min and at roomtemperature for 1.25 hrs, the mixture was acidified with 1.0N HCl andextracted with EtOAc. The extract was washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to a colorless,viscous oil (0.143 g, theory:0.130 g). TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.26.

To a solution of the crude acid (0.143 g, ca 0.395 mmol) and(3S)-3-(leucinyl)amino-4-oxobutanoic acid tert-butyl ester semicarbazone(see Example 1, Part B, 0.184 g, ca 0.41 mmol) inN-methylpyrrolidone(1.0 mL)-CH₂Cl₂(1.0 mL) at 0° C. (ice bath) undernitrogen was added hydroxybenzotriazole hydrate (0.077 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.120 g, 0.626 mmol). After stirring at 0° C. for 1.5 hrs and at roomtemperature for 3.5 hrs, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to give the title compound (0.270 g, 100%) as anoff-white foam. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.45. ¹H-NMR(300 MHz, CDCl₃)reveals that the product is a 82:18 mixture of 2′S (d, 0.91 ppm, 6.3 Hz;d, 0.95 ppm. 6.0 Hz) and 2′R (d, 0.56 ppm, 6.3 Hz; d, 0.67 ppm, 6.3 Hz)diastereomers due to racemization which occurred at some point in thesynthesis.

Part E:(3S,2′S)-3-[N-((2′-(1-Naphthyloxy)-4′-Carboxy)Butyryl)Leucinyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution of(3S,2′S)-3-[N-((2′-(1-naphthyloxy)-4′-carboxy)butyryl)leucinyl]amino-4-oxobutanoicacid semicarbazone di-tert-butyl ester (0.270 g, ca 0.395 mmol) inCH₂Cl₂(2.0 mL)-anisole(0.5 mL) at room temperature under nitrogen wasadded trifluoroacetic acid (2.5 mL). The resulting solution was stirredat room temperature for 4.5 hrs, evaporated to dryness and chased withCH₂Cl₂ and toluene-CH₂Cl₂ (1:1). The residue was triturated with EtOActo give the title compound (0.171 g, 80%) as an off-white solid.Evaporation of the mother liquor and trituration of the residue withEt₂O gave an additional 0.035 g of the title compound (total: 0.206 g,96%). TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.33. ¹H-NMR(300 MHz,CD₃OD) of the 1st crop of material indicates that the product is a 82:18mixture of 2′S (d, 0.88 ppm, 6.3 Hz; d, 0.95 ppm, 6.0 Hz) and 2′R (d,0.62 ppm, 6.6 Hz; d, 0.68 ppm, 6.3 Hz) diastereomers.

Part F:(3S,2′S)-3-[N-((2′-(1-Naphthyloxy)-4′-Carboxy)Butyryl)Leucinyl]Amino-4-OxobutanoicAcid

A suspension of(3S,2′S)-3-[N-((2′-(1-naphthyloxy)-4′-carboxy)butyryl)leucinyl]amino-4-oxobutanoicacid semicarbazone_(0.159 g, 0.29 mmol) in 37% aqueous formaldehyde(1.0mL)-acetic acid(1.0 mL)-methanol(3.0 mL) was stirred at room temperatureunder nitrogen for 18 hrs. The resulting clear solution was diluted withwater and extracted with EtOAc. The extract was washed with water andsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was taken up in tetrahydrofuran, filtered throughCelite and evaporated to dryness. The product was taken up in a smallamount of tetrahydrofuran and precipitated with Et₂O-hexane to give thetitle compound (0.121 g, 85%) as a white solid.TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.62. ¹H-NMR(300 MHz, CD₃OD)indicates that the product is a 78:22 mixture of 2′S (d, 0.88 ppm, 5.7Hz; d, 0.95 ppm, 6.0 Hz) and 2′R (d, 0.57 ppm, 6.6 Hz; 2d's, 0.659 ppm,6.6 Hz and 0.663 ppm, 6.6 Hz) diastereomers.

EXAMPLE 4

(3S,2′R)-3-[N-((2′-(1-Naphthyloxy)-4′-Carboxy)Butyryl)Leucinyl]Amino-4-OxobutanoicAcid

Starting with L-glutamic acid γ-benzyl ester following the method setforth in Example 3, Parts A through F, the title compound was similarlyprepared. ¹H-NMR(300 MHz, CD₃OD) indicates that the product is a 67:33mixture of 2′R (d, 0.57 ppm, 6.6 Hz-2d's, 0.659 ppm, 6.6 Hz and 0.663ppm, 6.6 Hz) and 2′S (d, 0.88 ppm, 5.7 Hz; d, 0.95 ppm, 6.0 Hz)diastereomers.

EXAMPLE 5

(3S)-3-[N-((1′-Carboxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Part A: (1-Carbomethoxy-2-Naphthyloxy)Acetic Acid

To a solution of 1-carbomethoxy-2-naphthol (0.382 g, 1.90 mmol) indimethylformamide (9.4 mL) at room temperature under nitrogen was addedtert-butyl bromoacetate (0.28 mL, 1.90 mmol) and powdered anhydrouspotassium carbonate (0.783 g, 5.7 mmol). After stirring at roomtemperature for 18 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water (2×) and saturated NaClsolution, dried over anhydrous sodium sulfate and evaporated to an oil.TLC(EtOAc-hexane; 1:10) Rf=0.18.

The crude product (ca 1.90 mmol) was taken up in CH₂Cl₂ (20 mL) andtreated with anisole (0.1 mL) and trifluoroacetic acid-water (9:1, 3.0mL) at room temperature under nitrogen. After stirring at roomtemperature for 16 hrs, the mixture was concentrated and chased withtoluene. Trituration of the residue with Et₂O-hexane gave the titlecompound (0.455 g, 92%) as a white solid.

Part B:(3S)-3-[N-((1′-Carbomethoxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of (1-carbomethoxy-2-naphthyloxy)acetic acid (0.260 g, 1.0mmol) in CH₂Cl₂(10 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (0.184 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.288 g, 1.50 mmol). After stirring for 15 min, the mixture was treatedwith (3S)-N-(valinyl)amino-4-oxobutanoic acid tert-butyl estersemicarbazone (0.329 g, 1.0 mmol, prepared by a method analogous to thatdescribed for N-(leucinyl)amino-4-oxobutanoic acid tert-butyl estersemicarbazone, see Example 1, Part B and Example 2, Part A) andN-methylmorpholine (0.13 mL, 1.2 mmol). After stirring at 0° C. for 2hrs and at room temperature for 16 hrs, the mixture was partitionedbetween EtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to give the title compound (0.571 g, 99%) as aviscous oil. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.63.

Part C:(3S)-3-[N-((1′-Carboxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of(3S)-3-[N-((1′-carbomethoxy-2′-naphthyloxy)acetyl)valinyl]amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.571 g, 1.0 mmol) in dioxane-water(3.0 mL, 3:1, v:v) at room temperature was added 1.0 N LiOH solution(1.1 mL, 1.1 mmol). After stirring at room temperature for 4 hrs, themixtuure was partitioned between EtOAc-5% KHSO₄. The organic phase waswashed with saturated NaCl solution, dried over anhydrous Na₂SO₄ andevaporated to dryness. Trituration with Et₂O-hexane gave the titlecompound (0.461 g, 83%) as a white solid. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.09.

Part D:(3S)-3-[N-((1′-Carboxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution of(3S)-3-[N-((1′-carboxy-2′-naphthyloxy)acetyl)valinyl]amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.279 g, 0.50 mmol) in CH₂Cl₂(5.0mL)-anisole(0.1 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (0.85 mL). The resulting clear solution was stirredat room temperature for 16 hr, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (0.241 g, 96%) as an off-white solid.

Part E:(3S)-3-[N-((1′-Carboxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

A solution of(3S)-3-[N-((1′-carboxy-2′-naphthyloxy)acetyl)valinyl]amino-4-oxobutanoicacid semicarbazone (0.100 g, 0.20 mmol) in MeOH-acetic acid-37% aqueousformaldhyde (4.0 mL, 3:1:1, v:v:v), was stirred at room temperatureunder nitrogen for 16 hrs. The mixture was concentrated, diluted withwater, frozen and lyophilized. The residue was taken up in methanol,filtered and evaporated to dryness. The residue was triturated with Et₂Oto give the title compound (0.070 g, 79%) as an off-white solid. MS(ES)for C₂₂H₂₄N₂O₈ (MW 444.44): positive 445(M+H); negative 443(M−H).

EXAMPLE 6

(3S)-3-[N-((2′-Carboxy-1′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Starting with 2-carbomethoxy-1-naphthol and following the generalmethods described in Example 5, Parts A through E, the title compoundwas also prepared. MS(ES) for C₂₂H₂₄N₂O₈ (MW 444.44): positive 445(M+H);negative 443(M−H).

EXAMPLE 7

(3S)-3-[N-((3′-Carboxy-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Starting with 3-carbomethoxy-2-naphthol and following the generalmethods described in Example 5, Parts A through E, the title compoundwas also prepared. MS(ES) for C₂₂H₂₄N₂O₈ (MW 444.44): positive 445(M+H),483(M+K); negative 443(M−H).

EXAMPLE 8

(3S)-3-[N-((1-Naphthylamino)Acetyl)Leucinyl]Amino-4-Oxobutanoic Acid

Part A: (1-Naphthylamino)Acetic Acid

To a solution of 1-aminonaphthalene (1.43 g, 10 mmol) and triethylamine(1.5 mL, 10.8 mmol) in dimethylformamide (5.0 mL) at room temperatureunder nitrogen was added methyl bromoacetate (1.5 mL, 15.8 mmol). Afterstirring at room temperature for 60 hrs, the mixture was partitionedbetween EtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to a purple oil (1.55 g). TLC(Et₂O-hexane; 2:3)major spot(UV and PMA) Rf=0.41 (1-aminonaphthalene Rf=0.33).

The crude methyl ester (1.55 g) was taken up in dioxane (10 mL) andtreated with 1.0N LiOH (12 mL, 12 mmol). After stirring at roomtemperature for 1 hr, the mixture was washed with Et₂O, the Et₂O washesdiscarded, and the aqueous phase acidified with 1.0N HCl (15 mL). Theresulting preciptate was collected by suction, washed with water andair-dried to give 1.35 g of crude product as a tan solid.Recrystallization fron EtOAc-hexane gave the title compound (1.03 g, 51%overall) as an off-white crystalline solid. TLC(MeOH-CH₂Cl₂; 1:9)Rf=0.16.

Part B: (3S)-3-[N-((1-Naphthylamino)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of (1-naphthylamino)acetic acid (0.076 g, 0.38 mmol) and(3S)-3-(leucinyl)amino-4-oxobutanoic acid tert-butyl ester semicarbazone(see Example 1, Part B, 0.180 g, ca 0.41 mmol) inN-methylpyrrolidone(1.0 mL)-CH₂Cl₂(1.0 mL) at 0° C. (ice bath) undernitrogen was added hydroxybenzotriazole hydrate (0.075 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.100 g, 0.52 mmol). After stirring at 0° C. for 3 hrs and at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The crude product was purified by flash chromatography elutingwith MeOH-CH₂Cl₂ (1:30 then 1:15) to give the title compound (0.201 g,100%) as a pale yellow foam. TLC(MeOH-CH₂Cl₂; 5:95) Rf=0.29.

Part C: (3S)-3-[N-((1-Naphthylamino)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution of(3S)-3-[N-((1-naphthylamino)acetyl)leucinyl]amino-4-oxobutanoic acidtert-butyl ester semicarbazone (0.201 g, 0.38 mmol) in CH₂Cl₂(2.0mL)-anisole(0.5 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (2.0 mL). The resulting solution was stirred atroom temperature for 5 hrs, evaporated to dryness and chased with CH₂Cl₂and toluene-CH₂Cl₂ (1:1). The resulting solid was triturated withCHCl₂-Et₂O to give the title compound (0.176 g, 98%) as a pale graysolid. TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.45.

Part D: (3S)-3-[N-((1-Nanhthylamino)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid

A solution of(3S)-3-[N-((1-naphthyloxy)acetyl)leucinyl]amino-4-oxobutanoic acidsemicarbazone (0.167 g, 0.36 mmol) in 37% aqueous formaldehyde(1.0mL)-acetic acid(1.0 mL)-methanol(3.0 mL) was stirred at room temperatureunder nitrogen for 4 hrs. The resulting solution was diluted with water,the resulting white preciptate collected by suction and washed withwater. The solid was air-dried, triturated with Et₂O and then dried invacuo to give the title compound (0.110 g, 75%) as a light gray solid.TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.54 (streaky spot).TLC(AcOH-MeOH-CH₂Cl₂; 1:1:8) Rf=0.25 (streaky spot).

EXAMPLE 9

(3S,2′RS)-3-[N-(2′-(1-Naphthylamino)Propionyl)Leucinyl]Amino-4-OxobutanoicAcid

Part A: 2-(1-Nanhthylamino)Propionic Acid

To a solution of 1-aminonaphthalene (1.43 g, 10 mmol) and triethylamine(1.5 mL, 10.8 mmol) in dimethylformamide (3.0 mL) at room temperatureunder nitrogen was added ethyl 2-bromopropionate (1.4 mL, 10.8 mmol).After stirring at 60° C. (bath temperature) 18 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to a brown oil. Purification of thecrude product by flash chromatography on silica gel eluting withEt₂O-hexane (5:95) to give ethyl 2-(1-naphthylamino)propionate (1.726 g,73%) as a white crystalline solid after trituration with cold hexane.TLC(Et₂O-hexane; 2:3) Rf=0.43.

The ethyl ester (1.644 g, 6.76 mmol) was taken up in dioxane (10 mL) andtreated with 1.0N LiOH (10 mL, 10 mmol). After stirring at roomtemperature for 1.5 hrs, the mixture was acidified with 1.0N HCl (12mL). The resulting preciptate was collected by suction, washed withwater and dried in vacuo to give the title compound (1.387 g, 95%) as awhite crystalline solid. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.38.

Part B:(3S,2′RS)-3-[N-(2′-(1-Naphthylamino)Propionyl)Leucinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of 2-(1-naphthylamino)propionic acid (0.081 g, 0.38 mmol)and (3S)-3-(leucinyl)amino-4-oxobutanoic acid tert-butyl estersemicarbazone (see Example 1, Part B, 0.180 g, ca 0.41 mmol) inN-methylpyrrolidone(1.0 mL)-CH₂Cl₂(1.0 mL) at 0° C. (ice bath) undernitrogen was added hydroxybenzotriazole hydrate (0.075 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.100 g, 0.52 mmol). After stirring at 0° C. for 2 hrs and at roomtemperature for 6 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness The crude product was purified by flash chromatography elutingwith MeOH-CH₂Cl₂ (1:30 then 1:15) to give the title compound (0.197 g,97%) as a white foam. TLC(MeOH-CH₂Cl₂; 5:95) Rf=0.35.

Part C:(3S,2′RS)-3-[N-(2′-(1-Nanhthylamino)Propionyl)Leucinyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution of(3S,2′RS)-3-[N-(2′-(1-naphthylamino)-propionyl)-leucinyl]-amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.184 g, 0.34 mmol) in CH₂Cl₂ (2.0mL)-anisole(0.5 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (2.5 mL). The resulting solution was stirred atroom temperature for 6.5 hrs, evaporated to dryness and chased withCH₂Cl₂ and toluene-CH₂Cl₂ (1:1). The resulting solid was triturated withCH₂Cl₂-Et₂O to give the title compound (0.148 g, 90%) as a pale graysolid. TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) two spots(diastereomers) Rf=0.36 and 0.39. TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) twospots (diastereomers) Rf=0.13 and 0.16.

Part D:(3S,2′RS)-3-[N-(2′-(1-Naphthylamino)Propionyl)Leucinyl]Amino-4-OxobutanoicAcid

A solution of(3S,2′RS)-3-[N-(2′-(1-naphthyloxy)propionyl)leucinyl]amino-4-oxobutanoicacid semicarbazone (0.138 g, 0.28 mmol) in 37% aqueous formaldehyde(0.5mL)-acetic acid(0.5 mL)-methanol(1.5 mL) was stirred at room temperatureunder nitrogen for 5.5 hrs. The resulting solution was diluted withwater (15 mL) and extracted with EtOAc. The extract was washed withwater and saturated NaCl solution, dried over anhydrous Na₂SO₄ andevaporated to dryness. The residue was taken up in methanol (0.5 mL) andapplied directly to a 3 mL Supelco™ LC-18 reverse phase extraction tubewhich had been pre-conditioned with water, and eluted successively with10 mL each of water, 30% MeOH-water, 60%. MeOH-water, 80% MeOH-water and90% MeOH-water. The product-containing fractions (TLC) were combined,concentrated and the resulting aqueous mixture extracted with EtOAc. Theextract was washed with saturated NaCl solution, dried over anhydrousNa₂SO₄ and evaporated to dryness. Trituration with EtOAc-Et₂O gave thetitle compound (0.098 g, 80%) as an off-white solid.TLC(EtOAc-pyridine-AcOH-H₂O; 60:20:5:10) Rf=0.50 (streaky spot).

EXAMPLE 10

(3S)-3-[N-((2,3-Dihydro-2,2-Dimethyl-Benzofuranyloxy)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid

Part A:(3S)-3-[N-(9-Fluorenylmethoxycabonyl)Leucinyl]Amino-4-Oxobutanoic Acidtert-Butyl EsterSemicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetyl)]Aminomethylpolystrene

Aminomethylpolystryene resin (10.0 g, 100–200 mesh, 0.71 meq/g) wasplaced in a 200 mL filter tube equipped with a vacuum stopcock and glassfrit and washed successively with CH₂Cl₂(50 mL)/dimethylformamide(50mL), diisopropylethylamine(5 mL)/dimethylformamide(30 mL),dimethylformamide (2×40 mL) and tetrahydrofuran (30 mL). The resin wassuspended in tetrahydrofuran(20 mL)/N-methylpyrolidinone(20 mL) withnitrogen agitation through the bottom of the frit and treated withdiiospropylethylamine (1.9 mL, 10.9 mmol) and(3S)-3-(9-fluorenylmethoxycabonyl)amino-4-oxobutanoic acid tert-butylester semicarbazonyl-4-[2′-(4-ethyl-phenoxyacetic acid)] (2.24 g, 3.56mmol). After all of the solid had dissolved (approx. 10 min), themixture was treated with pyBOP[benzotriazolyloxy-tris(N-pyrolidinyl)phosphonium hexafluorophosphate,2.78 g, 5.34 mmol) in one portion. After mixing by nitrogen agitationfor 3 hrs, the supernatant was removed by suction and the resin washedsuccesively with tetrahydrofuran (2×50 mL), dimethylformamide (3×50 mL)and CH₂Cl₂ (2×50 mL). Unreacted amine groups were capped by treatmentwith a mixture of acetic anhydride(10 mL)/dimethylformamide(30mL)/diisopropylethylamine(1.0 mL). After mixing by nitrogen agitationfor 1 hr, the supernatant was removed by suction and the resin washedwith dimethylformamide(4×50 mL).

The resin was treated with piperidine(10 mL)/dimethylformamide(40 mL)and mixed by nitrogen agitation for 45 min. The supernatant was removedby suction and the resin washed with dimethylformamide(4×50 mL) andtetrahydrofuran (50 mL).

The resin was suspended in tetrahydrofuran(20mL)/N-methylpyrolidinone(20 mL), treated withN-(9-fluorenylmethoxycabonyl)leucine (2.52 g, 7.12 mmol),diisopropylethylamine (3.8 mL, 21.8 mmol) and pyBOP (5.56 g, 10.7 mmol)and mixed by nitrogen agitation for 2.5 hrs. The supernatant was removedby suction and the resin washed successively with dimethylformamide(3×40 mL) and CH₂Cl₂ (3×40 mL), methanol (2×40 mL) and Et₂O (2×40 mL).The resin was dried in vacuo to give the title product (12.98 g,quanitative). Based on the starting semicarbazone-acid, the resinloading was calculated as approximately 0.27 meq/g.

Part B:(3S)-3-N-((2,3-Dihydro-2,2-Dimethyl-7-Benzofuranyloxy)Acetyl)Leucinyl]Amino-4-OxobutanoicAcid

An aliquot of the Part A resin (0.120 g, ca 0.032 mmol) was placed in a6 mL Supelco™ fitration tube equipped with a 20 μm polyethylene frit,treated with piperidine-dimethylformamide (1.0 mL, 1:4 v/v) and mixed onan orbital shaker for 1 hr. The supernatant was removed by suction andthe resin washed with dimethylformamide (4×1.0 mL) and CH₂Cl₂ (3×1.0mL). The resin was treated with 0.5M iPr₂NEt in N-methylpyrolidinone(0.40 mL, 0.20 mmol), (2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)aceticacid (0.026 g, 0.12 mmol) and 0.25M pyBOP in N-methylpyrolidinone (0.40mL, 0.10 mmol). The mixture was mixed on an orbital shaker under annitrogen atmosphere for 16 hrs. The supernatant was removed by suctionand the resin washed succesively with dimethylformamide (3×1.0 mL) andCH₂Cl₂ (3×1.0 mL), methanol (2×1.0 mL) and Et₂O (2×1.0 mL).

The resin was treated with 1.0 mL of CH₂Cl₂ and allowed to re-swell for15 min. The solvent was removed by suction and the resin treated withtrifluoroacetic acid-CH₂Cl₂-anisole (1.0 mL, 4:3:1 v/v/v). After mixingon an orbital shaker under nitrogen for 6 hrs, the supernatant wasremoved by suction and the resin washed with CH₂Cl₂ (4×1.0 mL). Theresin was treated with 37% aqueous formaldehyde-aceticacid-tetrahydrofuran-trifluoroacetic acid (1.0 mL, 1:1:5:0.025 v/v/v/v)and mixed on an orbital shaker under nitrogen for 4 hrs. The supernatantwas collected by suction, the resin washed with tetrahydrofuran (3×0.5mL). The combined filtrates were blown down under nitrogen. The residuewas taken up in methanol (0.5 mL), filtered and applied directly to a 3mL Supelco™ LC-18 reverse phase extraction tube which had beenpre-conditioned with water, and eluted successively with 3 mL each of10% MeOH-water, 30% MeOH-water, 60% MeOH-water and 90% MeOH-water. Theproduct-containing fractions (TLC) were combined and evaporated todryness to give the title compound (0.0084 g, 60%) as a colorless glass.TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.29.

EXAMPLES 11–52

Following the general procedure set forth in Example 10, Part B; thecompounds of Formula Ib (Examples 11 through 52) shown in Table 3 beloware also prepared. IC₅₀'s were determined by the method set forth inPrepartion 3A:

TABLE 3 Ex. mICE CPP32 MCH2 MCH3 MCH5 No. R¹ X n R² I₅₀(μM) I₅₀(μM)I₅₀(μM) I₅₀(μM) I₅₀(μM) 11 1-naphthyl CH₂ 0 H 1.86 1.59 4.19 8.78 12.212 1-naphthyl O 0 H 0.597 0.139 0.846 1.95 0.821 13 2-naphthyl O 0 H2.57 0.944 18.6 8.87 >10 14 1-naphthyl O 0 CH₃ 3.99 0.376 1.28 1.32 2.4315 6-Br-1-naphthyl O 0 CH₃ 6.84 4.81 13.8 32.4 29.1 16 1-naphthyl S 0 H2.75 0.195 1.43 1.74 7.42 17 2-naphthyl S 0 H 0.792 0.269 3.16 2.52 11.018 2-naphthyl CH₂ 1 H 1.80 2.76 14.5 18.2 >50 19 1-naphthyl C═O 1 H0.408 0.967 11.8 11.3 11.2 20 1-naphthyl C═O 1 CH₃ 4.55 9.88 24.9 29.83.25 21 2-naphthyl C═O 1 H 0.543 1.42 10.3 7.43 5.23 22 1-naphthyl O 1 H0.686 0.059 0.305 1.37 9.81 23 2-naphthyl O 1 H 1.32 0.910 5.90 9.6515.2 24 1-naphthyl S 1 H 0.563 0.412 2.72 3.60 16.3 25 2-naphthyl S 1 H0.611 0.837 1.62 5.89 15.0 26 2-Me-1-naphthyl O 0 H 0.843 0.375 32.44.16 4.14 27 4-MeO-1-naphthyl O 0 H 0.831 0.263 22.6 4.08 1.45 284-Cl-1-naphthyl O 0 H 0.429 0.231 12.0 3.38 1.69 29 2,4-diCl-1-naphthylO 0 H 0.141 0.357 21.4 3.61 3.04 30 1-isoquinolinyl O 0 H 44.2 1.57 >5034.7 >50 31 4-quinolinyl O 0 H 35.3 0.232 >50 4.57 >50 32 5-quinolinyl O0 H 5.25 0.412 >50 3.85 4.02 33 5-isoquinolinyl O 0 H 5.14 0.407 42.73.48 3.64 34 8-quinolinyl O 0 H 13.7 0.147 12.5 1.51 2.24 35 phenyl CH₂0 H >10 9.74 ND >10 >10 36 phenyl O 0 CH₃ 20.4 1.77 >10 8.27 >10 37phenyl O 1 H 9.42 0.419 >50 6.04 >10 38 phenyl O 0 H >103.40 >50 >10 >10 39 2-biphenyl O 0 H 0.636 0.095 0.717 2.02 1.71 403-biphenyl O 0 H 1.10 0.311 14.5 3.75 3.86 41 4-biphenyl O 0 H 1.900.763 20.5 12.0 7.53 42 (2-benzyl)phenyl O 0 H 0.521 0.490 10.1 3.366.05 43 (4-benzyl)phenyl O 0 H 1.80 0.346 18.9 4.41 4.72 44(4-phenoxy)phenyl O 0 H 2.21 0.545 21.2 6.82 9.28 45 (2-benzyloxy)phenylO 0 H 2.40 0.222 9.75 2.20 4.34 46 (4-benzyloxy)phenyl O 0 H 2.51 0.57033.4 7.25 8.60 47 (2-cyclo-pentyl)- O 0 H 0.538 0.197 3.37 1.49 1.86phenyl 48 (4-cyclo-pentyl)- O 0 H 2.20 0.319 51.2 5.23 5.90 phenyl 49[2-(1-adamantanyl)- O 0 H 1.43 0.474 5.86 2.79 3.87 4-Me]phenyl 504-(1-adamantanyl)- O 0 H 1.83 0.528 32.5 8.24 4.35 phenyl 515,6,7,8-tetrahydro-1- O 0 H 1.81 0.324 11.8 2.74 1.75 naphthyl 525,6,7,8-tetrahydro-2- O 0 H 2.57 0.162 28.6 2.31 4.95 naphthyl

EXAMPLE 53

(3RS)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-Fluoro-4-OxopentanoicAcid

Part A:(3RS,4RS)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid tert-Butyl Ester

To a solution of (N-benzyloxycarbonyl)valine (0.332 g, 1.32 mmol) inCH₂Cl₂(7.0 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (0.219 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.317 g, 1.65 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with (3RS,4RS)-3-amino-5-fluoro-4-hydroxypentanoic acid,tert-butyl ester (0.228 g, 1.1 mmol, prepared as described inTetrahedron Letters 1994, 35, 9693–9696) and the reacton allowed to warmto room temperature. After stirring at room temperature for 24 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography eluting with EtOAc-hexane(1:1) to give the title compound (0.423 g, 87%) as colorless glass.TLC(MeOH-CH₂Cl₂; 5:95) Rf=0.17.

Part B: (3RS,4RS)-3-(Valinyl)Amino-5-Fluoro-4-Hydroxypentanoic Acid,tert-Butyl Ester

To a solution of(3RS,4RS)-3-[(N-benzyloxycarbonyl)valinyl]amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (1.00 g, 2.30 mmol) in EtOH (130 mL) was added10% Pd—C (0.120 g) and resulting mixture stirred under a hydrogenatmosphere (balloon) for 1 hr. The mixture was filtered through Celitewashing the filter cake with CH₂Cl₂ and the combined filtratesevaporated to dryness. The residue was chased with CH₂Cl₂ to give thetitle product (0.707 g, quantitative) as a colorless oil.TLC(MeOH-CH₂Cl₂; 1;9) Rf=0.50.

Part C:(3RS,4RS)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester

To a solution of (1-naphthyloxy)acetic acid (0.202 g, 1.0 mmol) in indimethylformamide(4.0 mL)-CH₂Cl₂ (6.0 mL) at 0° C. (ice bath) undernitrogen was added hydroxybenzotriazole hydrate (0.168 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.249 g, 1.3 mmol). After stirring for 10 min, the mixture was treatedwith a solution of(3RS,4RS)-3-(valinyl)amino-5-fluoro-4-hydroxypentanoic acid, tert-butylester (0.319 g, 1.04 mmol) in CH₂Cl₂(8.0 mL). After stirring at 0° C.for 1 hr and at room temperature for 3 hrs, the mixture was partitionedbetween EtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (3:2) to give thetitle compound (0.307 g, 63%) as a white solid. TLC(MeOH-CH₂Cl₂; 1:9)Rf=0.69.

Part D:(3RS)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-Fluoro-4-OxopentanoicAcid, tert-Butyl Ester

To a solution of(3RS,4RS)-3-[N-((1-naphthyloxy)acetyl)valinyl]amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (0.163 g, 0.315 mmol) and N-methylmorpholineN-oxide (0.144 g, 0.98 mmol) in CH₂Cl₂ (5.0 mL) at room temperature wasadded activated 4 Å molecular sieves. After stirring at room temperaturefor 20 min, the mixture was treated with tetra(n-propyl)ammoniumperruthenate (0.011 g). After stirring at room temperature for 3.5 hrs,the mixture through Celite and evaporated to dryness. The residue waspurified by flash chromatography on silica gel eluting with EtOAc-hexane(3:4) to give the title compound (0.124 g, 40%) as a pale yellow oil.TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.71. ¹H NMR (CDCl₃): δ 8.27–8.23 (m, 1H),7.86–7.83 (m, 1H), 7.59–7.51 (m, 3H), 7.42–7.36 (m, 1H), 7.23–7.19 (m,1H), 7.05–6.95 (m, 1H), 6.84 (d, 1H, J=7.7 Hz), 5.26–4.97 (m, 2H),4.93–4.89 (m, 1H), 4.76 (s, 2H), 4.45–4.35 (m, 1H), 3.05–2.76 (m, 2H),1.42 (d, 9H, J=4.1 Hz), 1.02–0.87 (m, 6H).

Part E:(3RS)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-Fluoro-4-OxopentanoicAcid

To a solution of(3RS)-3-[N-((1-naphthyloxy)acetyl)valinyl]amino-5-fluoro-4-oxopentanoicacid, tert-butyl ester (0.113 g, 0.23 mmol) in CH₂Cl₂(2.0mL)-anisole(0.5 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (1.0 mL). The resulting clear solution was stirredat room temperature for 1 hr, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was purified by flash chromatographyon silica gel eluting with AcOH-MeOH-CH₂Cl₂ (0.5:2:100) to give thetitle compound (0.069 g, 69%) as a white solid. TLC(AcOH-MeOH-CH₂Cl₂;1:1:20) Rf=0.38. MS(ES) for C₂₂H₂₅FN₂O₆ (MW 432.45): positive 433(M+NH);negative 431(M−H). ¹H NMR (CD₃OD): δ 8.32–8.29 (m, 1H), 7.82–7.79 (m,1H), 7.49–7.46 (m, 3H), 7.38–7.32 (m, 1H), 6.88 (d, 1H, J=7.7 Hz),4.78–4.73 (m, 2H), 4.55–4.26 (m, 2H), 2.82–2.76 (m, 2H), 2.16–2.03 (m,1H), 0.94–0.85 (m, 6H).

EXAMPLES 54–56

Starting with (3RS,4RS)-3-(valinyl)amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (see Example 53, Part B) and following themethods described in Example 53, Parts C through E, the compounds shownbelow in Table 4 were also prepared:

TABLE 4

MS(ES) Ex. R¹ X n R² Formula MW pos. neg. 54 2-naphthyl O 0 HC₂₂H₂₅FN₂O₆ 432.45 433(M + H) 431(M − H) 455(M + Na) 545(M + TFA)471(M + K) 55 1-naphthyl O 1 H C₂₃H₂₇FN₂O₆ 446.47 447(M + H) 445(M − H)489(M + Na) 559(M + TFA) 56 (2-Ph)Ph O 0 H C₂₄H₂₇FN₂O₆ 458.49 481(M +Na) 457(M − H) 497(M + K) 571(M + TFA)

EXAMPLE 57

(3RS)-3-[N-((2-Phenylphenoxy)Acetyl)Leucinyl]Amino-5-Fluoro-4-OxopentanoicAcid

Part A:(3RS,4RS)-3-[(N-Benzaloxycarbonyl)Leucinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester

To a solution of (3RS,4RS)-3-amino-5-fluoro-4-hydroxypentanoic acid,tert-butyl ester (0.230 g, 1.1 mmol) in CH₂Cl₂ (2.0 mL) at roomtemperature under nitrogen was added (N-benzyloxycarbonyl)leucine,N-hydroxysuccinimde ester (0.402 g, 1.1 mmol). After stirring at roomtemperature for 16 hrs, the mixture was evaporated to dryness and theresidue purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:2) to give the title compound (0.332 g, 66%) as acolorless, viscous oil. TLC(EtOAc-hexane; 2:1) Rf=0.51.

Part B: (3RS,4RS)-3-(Leucinyl)Amino-5-Fluoro-4-Hydroxypentanoic Acid,tert-Butyl Ester, p-Toluenesulfonate Salt

To a solution of(3RS,4RS)-3-[(N-benzyloxycarbonyl)leucinyl]amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (0.332 g, 0.734 mmol) in MeOH (100 mL) was addedp-toluenesulsufonic acid hydrate (0.140 g, 0.737 mmol) and 10% Pd—C(0.033 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 2 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated todryness. The residue was chased with CH₂Cl₂ to give the title product(0.371 g) as a colorless foam.

Part C:(3RS)-3-[N-((2-Phenylphenoxy)Acetyl)Leucinyl]Amino-5-Fluoro-4-OxopentanoicAcid

Starting with (3RS,4RS)-3-(leucinyl)amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester, p-toluenesulfonate salt and following themethods described in Example 53, Parts C through E utilizing(2-phenylphenoxy)acetic acid in place of (1-naphthyloxy)acetic acid inPart C, gave the title compound as a white solid. MS(ES) for C₂₅H₂₉FN₂O₆(MW 472.51): positive 495(M+Na), 511(M+K); negative 471(M−H),585(M+TFA).

EXAMPLE 58

(3RS)-3-[N-(1′-Naphthyloxy)Propionyl)Leucinyl]Amino-5-Fluoro-4-OxopentanoicAcid

Starting with (3RS,4RS)-3-(leucinyl)amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester, p-toluenesulfonate salt and following themethods described in Example 53, Parts C through E utilizing3-(1′-naphthyloxy)propionic acid in place of (1-naphthyloxy)acetic acidin Part C, gave the title compound as a white solid. MS(ES) forC₂₄H₂₉FN₂O₆ (MW 460–50): positive 479(M+Na); negative 569(M+TFA).

EXAMPLE 59

(S,3RS)-3-[N-(1-Naphthyloxy)Acetyl)Homoprolinyl]Amino-5-Fluoro-4-OxopentanoicAcid

Following the general methods described in Example 53, Parts A throughE, and utilizing N-(benzyloxycarbonyl)-homoproline in place ofN-(benzyloxycarbonyl)valine in Part A, the title compound was alsoprepared. TLC(CH₂Cl₂/MeOH/AcOH, 20:1:1): Rf=0.50. ¹H NMR (CD₃OD): δ8.34–8.31 (m, 1H), 7.82–7.79 (m, 1H), 7.49–7.34 (m, 4H), 6.91–6.89 (m,1H), 5.20–3.93 (m, 6H), 3.06–2.50 (m, 2H), 2.36–2.14 (m, 2H), 1.80–1.22(m, 6H). MS(ES) for C₂₃H₂₅FN₂O₆ (MW 444.46): positive 445(M+H): negative443(M−H).

EXAMPLE 60

(2′S,3RS)-3-[N-(1-Naphthyloxy)Acetyl)-1,2,3,4-Tetrahydroisoquinoline-2′-Carbonyl]Amino-5-Fluoro-4-OxopentanoicAcid

Following the general methods described in Example 53, Parts A throughE, and utilizing(2S)-N-(benzyloxycarbonyl)-1,2,3,4-tetrahydroisoquinoline-2-carboxylicacid in place of N-(benzyloxycarbonyl)valine in Part A, the titlecompound was also prepared. MS(ES) for C₂₇H₂₅FN₂O₆ (MW 492.50): positive493(M+H); negative 491(M−H).

EXAMPLE 61

(2′S,3RS)-3-[N-((1-Naphthyloxy)Acetyl)Indoline-2′-Carbonyl]Amino-5-Fluoro-4-OxopentanoicAcid

Part A: (2S)-N-[(1-Naphthyloxy)Acetyl]Indoline-2′-Carboxylic Acid,Methyl Ester

To a solution of (1-naphthyloxy)acetic acid (1.119 g, 5.53 mmol) inether (30 mL) at 0° C. was treated with phosphorus pentachloride (1.267g, 6.08 mmol). After stirring at 0° C. for 20 min and at roomtemperature for 30 min, the mixture was evaporated to dyness and theresidue chased with toluene (2×) to give a light-yellow oil. The crudeacid chloride was taken up in toluene (10 mL) and added to a vigorouslystirring mixture of methyl (S)-indoline-2-carboxylate hydrochloride(1.182 g, 5.53 mmol) in toluene (10 mL)/aqueous NaHCO₃ solution (2.1 gin 18 mL of H₂O) under N₂ at 0° C. The mixture was stirred for 30 minthen partitioned between EtOAc and 5% KHSO₄. The organic phase waswashed with 5% KHSO₄, sat'd NaHCO₃ (2×) and saturated NaCl solutions,dried (Na₂SO₄), and evaporated to dryness to give the title compound(1.986 g, 99%) as a white foam.

Part B: (2S)-N-[(1-Naphthyloxy)Acetyl]Indoline-2-Carboxylic Acid

To a solution of (2S)-N-[(1-naphthoxy)acetyl]indoline-2-carboxylic acidmethyl ester (1.0 g, 2.77 mmol) in tetrahydrofuran (3.3 mL) at 0° C. wasadded 1.0 N LiOH solution (3.3 mL, 3.3 mmol). After stirring at 0° C.for 2 hours the mixture was concentrated, diluted with water, acidifiedto pH 3, and extracted with EtOAc. The EtOAc extract was washed withsaturated NaCl, dried (Na₂SO₄), and evaporated to give the titlecompound (0.918 g, 96%) as an off-white solid. ¹H NMR (CD₃OD): δ8.36–8.33 (m, 1H), 8.15 (d, 1H, J=7.8 Hz), 7.81–7.78 (m, 1H), 7.49–7.18(m, 7H), 7.10–7.04 (m, 1H), 6.92 (d, 1H, J=7.5 Hz), 5.32–4.94 (m, 5H),3.69–3.34 (m, 2H).

Part C:(2′S,3RS,4RS)-N-[((1-Naphthyloxy)Acetyl)Indoline-2′-Carbonyl]Amino-5-Fluoro-4-HydroxypentanoicAcid t-Butyl Ester

To a solution of (2S)-N-[(1-naphthyloxy)acetyl]-indoline-2-carboxylicacid (0.278 g, 0.8 mmol) in CH₂Cl₂ (2.0 mL)-dimethylformamide (0.5 mL)at 0° C. under nitrogen was added hydroxybenzotriazole hydrate (0.129 g)followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (0.184 g, 0.96 mmol). After stirring at 0° C. for 10 min,a solution of (3RS,4RS)-3-amino-5-fluoro-4-hydroxypentanoic acid,tert-butyl ester (0.166 g, 0.8 mmol) in CH₂Cl₂ (3.0 mL) was added. Afterstirring at 0° C. for 1 hr and at room temperature for 3 hrs, thereaction mixture was partitioned between EtOAc and 5% KHSO₄. The organicphase was washed with 5% KHSO₄, saturated NaHCO₃ (2×) and saturated NaClsolutions, dried (Na₂SO₄). and evaporated to dryness to give the crudetitle compound (255 mg) as an off-white solid. TLC(CH₂Cl₂-MeOH, 9:1):Rf=0.60.

Part D:(2′S,3RS)-N-[((1-Naphthyloxy)Acetyl)Indoline-2′-Carbonyl]Amino-5-Fluoro-4-OxopentanoicAcid t-Butyl Ester

To a solution of 2.0 M oxalyl chloride-CH₂Cl₂ (0.3 mL, 0.6 mmol) at −78°C. under nitrogen was added dimethylsulfoxide (0.09 mL, 1.2 mmol). Afterstirring at −78° C. for 10 min, a solution of(2′S,3RS,4RS)-N-[((1-naphthyloxy)acetyl)indoline-2′-carbonyl]amino-5-fluoro-4-hydroxypentanoicacid t-butyl ester (0.255 g, 0.48 mmol) in dry CH₂Cl₂ (3.0 mL) was addeddropwise. After stirring at −78° C. for 15 min, triethylamine (0.27 mL,2.5 mmol) was added dropwise, the mixture stirred for 10 min, thenallowed to warm to room temperature. After an additional 1 hr, themixture was partitioned between EtOAc and 5% KHSO₄. The organic phasewas washed with 5% KHSO₄ and saturated NaCl solutions, dried (Na₂SO₄),and evaporated to a yellow oil. The crude product was purified by flashchromatography on silica gel eluting with EtOAc/hexane (1:2) to give thetitle compound (0.214 g, 83%) as a pale yellow solid. TLC(EtOAc/hexane,1:1): Rf=0.50.

Part E:(2′S,3RS)-N-[((1-Naphthyloxy)Acetyl)Indoline-2′-Carbonyl]Amino-5-Fluoro-4-OxopentanoicAcid

To a solution of(2′S,3RS)-N-[((1-naphthyloxy)acetyl)indoline-2′-carbonyl]amino-5-fluoro-4-oxopentanoicacid t-butyl ester (0.107 g, 0.20 mmol) in anisole (0.2 mL)-CH₂Cl₂ (2.0mL) at room temperature under nitrogen was added trifluoroacetic acid(1.0 mL). After stirring at room temperature for 1.5 hrs, the mixturewas concentrated then chased with CH₂Cl₂ and toluene. The reside wastrituated with ether-hexane to give the title ccompound (0.065 g, 68%)as an off-white solid. ¹H NMR (CD₃OD): δ 8.32–8.17 (m, 2H), 7.81–7.79(m, 1H), 7.54–6.80 (m, 8H), 5.38–4.29 (m, 6H), 3.25–2.32 (m, 4H). MS(ES)for C₂₆H₂₃FN₂O₆ (MW 478.48): positive 479 (M+H); negative 477 (M−H).

EXAMPLE 62

(3S)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-(1′,2′,3′-Benzotriazin-4′(3H)-on-3′-yloxy)-4-OxopentanoicAcid

Part A: [(N-Benzyloxycarbonyl)Valinyl]Aspartic Acid, β-tert-Butyl,α-Methyl Ester

To a solution of (N-benzyloxycarbonyl)valine (2.10 g, 8.36 mmol) inCH₂Cl₂(20 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (1.74 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(2.40 g, 12.5 mmol). After stirring at 0° C. for 10 min, the mixture wastreated with aspartic acid, β-tert-butyl, α-methyl ester hydrochloride(2.00 g, 8.34 mmol) and N-methylmorpholine 1.1 mL, 10 mmol), and thereaction allowed to warm to room temperature. After stirring at roomtemperature for 2.5 hrs, the mixture was concentrated and the residuepartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to give the title compound (3.55 g,97%) as a white solid after tituration with Et₂O-hexane.TLC(EtOAc-hexane; 1:1) Rf=0.48.

Part B: N-(Valinyl)Aspartic Acid, β-tert-Butyl, α-Methyl Ester

To a solution of [(N-benzyloxycarbonyl)valinyl]aspartic acid,β-tert-butyl, α-methyl ester (2.14 g, 4.90 mmol) in EtOH (200 mL) wasadded 10% Pd—C (0.21 g) and resulting mixture stirred under a hydrogenatmosphere (balloon) for 2 hrs. The mixture was filtered through Celitewashing the filter cake with CH₂Cl₂ and the combined filtratesevaporated to dryness. The residue was chased with CH₂Cl₂ to give thetitle product (1.48 g, quantitative) as a viscous oil. The crude productwas used immediately for the next step.

Part C: [N-((1-Naphthyloxy)Acetyl)Valinyl]Aspartic Acid, β-tert-Butyl,α-Methyl Ester

To a solution of (1-naphthyloxy)acetic acid (0.936 g, 4.90 mmol) inCH₂Cl₂(45 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (0.851 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(1.33 g, 6.94 mmol). After stirring for 15 min. the mixture was treatedwith N-(valinyl)aspartic acid, β-tert-butyl, α-methyl ester (1.48 g, ca4.90 mmol) and N-methylmorpholine (0.61 mL, 5.55 mmol). After stirringat 0° C. for 2 hrs and at room temperature for 16 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to dryness. The residue waspurified by flash chromatography on silica gel eluting with EtOAc-hexane(1:2) to give the title compound (1.89 g, 79%) as a viscous oil.TLC(EtOAc-hexane; 1:1) Rf=0.57.

Part D: [N-((1-Naphthyloxy)Acetyl)Valinyl]Aspartic Acid, β-tert-ButylEster

To a solution of [N-((1-naphthyloxy)acetyl)valinyl]aspartic acid,β-tert-butyl, α-methyl ester (1.88 g, 3.87 mmol) in dioxane (9.0mL)-water (3.0 mL) was added 1.0 N LiOH solution (4.3 mL, 4.3 mmol).After stirring at room temperature for 1 hr, the mixture was acidifiedwith 1.0 N HCl and extracted with EtOAc. The extract was washed withsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated togive the title compound (1.82 g, quantitative) as a white solid.TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.65.

Part E:(3S)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-Bromo-4-OxopentanoicAcid tert-Butyl Ester

To a solution of [N-((1-naphthyloxy)acetyl)valinyl]aspartic acid,β-tert-butyl ester (3.96 g, 8.40 mmol) and N-methylmorpholine (1.48 mL,13.5 mmol) in tetrahydrofuran (37 mL) at −10° C. (NaCl/ice bath) undernitrogen was added isobutyl chloroformate (1.63 mL, 12.6 mmol). Afterstirring at −10° C. for 0.5 hrs, the mixture was filtered into anotherice-cooled flask and the filter cake washed with cold tetrahydrofuran(approx. 15 mL). The resulting mixed anhydride solution was treated at−10° C. with excess diazomethane/Et₂O solution (prepared from 3.09 g, 21mmol of 1-methyl-3-nitro-1-nitrosoguanidine, 15 mL 40% KOH/28 mL Et₂O).After stirring at −10° C. for 30 min and at room temperature for 15 min,the mixture was cooled to 0° C. (ice bath) and treated with 48% aqueousHBr (19.0 mL, 170 mmol). Gas evolution was observed. After 15 min, themixture was partitioned between EtOAc-saturated NaHCO₃, the organicphase washed with saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. Trituration of the residue with Et₂O gave the titlecompound (3.29 g, 71%) as a white solid. TLC(EtOAc-hexane; 1:1) Rf=0.51.¹H NMR (CDCl₃): δ 8.26–8.22 (m, 1H), 7.86–7.83 (m, 1H), 7.59–7.51 (m,3H), 7.41–7.36 (m, 1H), 7.27–7.20 (m, 2H), 6.83 (d, 1H, J=7.8 Hz),5.00–4.95 (m, 1H), 4.76 (s, 2H), 4.48–4.43 (m, 1H), 4.12 (s, 2H),2.95–2.74 (dd, 2H), 2.26–2.19 (m, 1H), 1.41 (s, 9H), 0.99 (d, 3H, J=6.9Hz), 0.92 (d, 3H, J=6.9 Hz).

Part F:(3S)-3-[N-((1-Naphthyloxy)Acetyl)Valinyl]Amino-5-(1′,2′,3′-Benzotriazin-4′(3H)-on-3′-yloxy)-4-OxopentanoicAcid, tert-Butyl Ester

To a solution of(3S)-3-[N-((1-naphthyloxy)acetyl)valinyl]amino-5-bromo-4-oxopentanoicacid tert-butyl ester (0.165 g, 0.30 mmol) and3-hydroxy-1,2,3-benzotriazin-4(3H)-one (0.059 g, 0.36 mmol) indimethylformamide (2.0 mL) at room temperature under nitrogen was addedpotassium fluoride (0.061 g, 1.05 mmol). After stirring at roomtemperature for 5 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. Trituration of the residue with Et₂O-hexane gave the titlecompound (0.171 g, 90%) as pale yellow solid. TLC(MeOH-CH₂Cl₂; 1:9)Rf=0.40.

Part G:(3S)-3-[N-((1-Naphthyloxy)Acetyl)Valiny]Amino-5-(1′,2′,3′-Benzotriazin-4′(3H)-on-3′-yloxy)-4-OxopentanoicAcid

To a solution of(3S)-3-[N-((1-naphthyloxy)acetyl)valinyl]amino-5-(1′,2′,3′-benzotriazin-4′(3H)-on-3′-yloxy)-4-oxopentanoicacid, tert-butyl ester (0.143 g, 0.23 mmol) in CH₂Cl₂(2.0mL)-anisole(0.2 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (1.0 mL). The resulting clear solution was stirredat room temperature for 2 hr. evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O-hexane togive the title compound (0.099 g, 76%) as an off-white solid. ¹H NMR(CD₃OD): δ 8.33–7.24 (m, 10OH), 6.92–6.77 (m, 1H), 5.38–5.27 (m, 1H),4.80–4.31 (m, 5H), 3.08–2.60 (m, 2H), 2.18–2.04 (m, 1H), 1.11–0.83 (m,6H). MS(ES) for C₂₉H₂₉N₅O₈ (MW 575.58): positive 576(M+H); negative574(M−H).

EXAMPLES 63–149

Starting with(3S)-3-[N-((1-naphthyloxy)acetyl)valinyl]amino-5-bromo-4-oxopentanoicacid tert-butyl ester (see Example 62, Part E) and following the methodsdescribed in Example 62, Parts F through G, the compounds shown below inTable 5 were also prepared:

TABLE 5

MS(ES) Ex. B Formula MW pos. neg. 63 CH₂OCO(2,6-diCl-Ph) C₂₉H₂₈Cl₂N₂O₈603.45 603/605 601/603 (M + H) (M − H) 64 CH₂OPh C₂₈H₃₀N₂O₇ 506.55507(M + H) 505(M − H) 529(M + Na) 545(M + K) 65 CH₂O(2-F—Ph) C₂₈H₂₉FN₂O₇524.54 525(M + H) 523(M − H) 66 CH₂O(3-F—Ph) C₂₈H₂₉FN₂O₇ 524.54 525(M +H) 523(M − H) 67 CH₂O(4-F—Ph) C₂₈H₂₉FN₂O₇ 524.54 547(M + Na) 523(M − H)68 CH₂O(2,3-diF-Ph) C₂₈H₂₉F₂N₂O₇ 542.54 543(M + H) 541(M − H) 565(M +Na) 655(M + TFA) 69 CH₂O(2,4-diF-Ph) C₂₈H₂₉F₂N₂O₇ 542.54 543(M + H)541(M − H) 565(M + Na) 581(M + K) 70 CH₂O(2,5-diF-Ph) C₂₈H₂₉F₂N₂O₇542.54 543(M + H) 541(M − H) 565(M + Na) 581(M + K) 71 CH₂O(2,6-diF-Ph)C₂₈H₂₉F₂N₂O₇ 542.54 543(M + H) 541(M − H) 565(M + Na) 72CH₂O(3,4-diF-Ph) C₂₈H₂₉F₂N₂O₇ 542.54 543(M + H) 541(M − H) 581(M + K) 73CH₂O(3,5-diF-Ph) C₂₈H₂₉F₂N₂O₇ 542.54 543(M + H) 541(M − H) 565(M + Na)581(M + K) 74 CH₂O(2,3,4-triF-Ph) C₂₈H₂₇F₃N₂O₇ 560.53 561(M + H) 559(M −H) 583(M + Na) 599(M + K) 75 CH₂O(2,3,5-triF-Ph) C₂₈H₂₇F₃N₂O₇ 560.53561(M + H) 559(M − H) 583(M + Na) 673(M + TFA) 599(M + K) 76CH₂O(2,3,6-triF-Ph) C₂₈H₂₇F₃N₂O₇ 560.53 561(M + H) 559(M − H) 583(M +Na) 673(M + TFA) 599(M + K) 77 CH₂O(2,4,5-triF-Ph) C₂₈H₂₇F₃N₂O₇ 560.53561(M + H) 559(M − H) 583(M + Na) 599(M + K) 78 CH₂O(2,4,6-triF-Ph)C₂₈H₂₇F₃N₂O₇ 560.53 561(M + H) 559(M − H) 583(M + Na) 79CH₂O(2,3,5,6-tetraF-Ph) C₂₈H₂₆F₄N₂O₇ 578.52 579(M + H) 577(M − H)601(M + Na) 617(M + K) 80 CH₂O(2,3,4,5,6-pentaF-Ph) C₂₈H₂₅F₄N₂O₇ 596.51619(M + Na) 595(M − H) 81 CH₂O(2-CF₃—Ph) C₂₉H₂₉F₃N₂O₇ 574.55 597(M + Na)573(M − H) 82 CH₂O(3-CF₃—Ph) C₂₉H₂₉F₃N₂O₇ 574.55 597(M + Na) 573(M − H)83 CH₂O(4-CF₃—Ph) C₂₉H₂₉F₃N₂O₇ 574.55 597(M + Na) 573(M − H) 84CH₂O(3,5-diCF₃—Ph) C₃₀H₂₈F₆N₂O₇ 642.55 643(M + H) 641 (M − H) 665(M +Na) 681(M + K) 85 CH₂O(2-F,3-CF₃—Ph) C₂₉H₂₈F₄N₂O₇ 592.54 593(M + H) 591(M − H) 615(M + Na) 631(M + K) 86 CH₂O(2,6-diCl-Ph) C₂₈H₂₈Cl₂N₂O₇ 575.44575/577 573/575 (M + H) (M − H) 87 CH₂O(2-NO₂—Ph) C₂₈H₂₉N₃O₉ 551.55552(M + H) 550(M − H) 574(M + Na) 590(M + K) 88 CH₂O(4-NO₂—Ph)C₂₈H₂₉N₃O₉ 551.55 552(M + H) 550(M − H) 574(M + Na) 89CH₂O(2-F,4-NO₂—Ph) C₂₈H₂₈FN₃O₉ 569.54 570(M + H) 568(M − H) 592(M + Na)90 CH₂O(4-CN—Ph) C₂₉H₂₉N₃O₇ 531.56 554(M + Na) 530(M − H) 91CH₂O(4-CF₃O—Ph) C₂₉H₂₉F₃N₂O₈ 590.55 591(M + H) 589(M − H) 703(M + TFA)92 CH₂O(4-H₂NCO—Ph) C₂₉H₃₁N₃O₈ 549.58 550(M + H) 548(M − H) 572(M + Na)662(M + TFA) 93 CH₂O(4-PhCO—Ph) C₃₅H₃₄N₂O₈ 610.66 611(M + H) 609(M − H)633(M + Na) 94 CH₂O(4-Ph—Ph) C₃₄H₃₄N₂O₇ 582.65 583(M + H) 581(M − H)605(M + Na) 695(M + TFA) 621(M + K) 95 CH₂O(4-C₆F₅-2,3,5,6-tetraF-Ph)C₃₄H₂₅F₉N₂O₇ 744.57 745(M + H) 743(M − H) 767(M + Na) 783(M + K) 96CH₂O(4-PhO—Ph) C₃₄H₃₄N₂O₈ 598.65 599(M + H) 597(M − H) 621(M + Na) 97CH₂O[4-(4′-CF₃—PhO)Ph] C₃₅H₃₃F₃N₂O₈ 666.65 667(M + H) 665(M − H) 689(M +Na) 98 CH₂O(3-AcNH—Ph) C₃₀H₃₃N₃O₈ 563.61 564(M + H) 562(M − H) 586(M +Na) 99 CH₂O(3,4-OCOS—Ph) C₂₉H₂₈N₂O₉S 580.61 581(M + H) 693(M + TFA)603(M + Na) 619(M + K) 100 CH₂O(2-pyridinyl) C₂₇H₂₉N₃O₇ 507.54 508(M +H) 506(M − H) 101 CH₂O(4,5-diCl-3-pyridazinyl) C₂₆H₂₆Cl₂N₄O₇ 577.42577/579 575/577 (M + H) (M − H) 689/691 (M + TFA) 102 CH₂O(2-naphthyl)C₃₂H₃₂N₂O₇ 556.61 557(M + H) 555(M − H) 103 CH₂OPOPh₂ C₃₄H₃₅N₂O₈P 630.63631 (M + H) 629(M − H) 653(M + Na) 104 CH₂OPO(Me)Ph C₂₉H₃₃N₂O₈P 568.56569(M + H) 567(M − H) 105 CH₂OPOMe₂ C₂₄H₃₁N₂O₈P 506.49 529(M + Na) 505(M− H) 106 CH₂OPO(n-hexyl)Ph C₃₄H₄₃N₂O₈P 638.28 639(M + H) 637(M − H)661(M + Na) 751(M + TFA) 677(M + K) 107 CH₂OPO(PhCH₂)Ph C₃₅H₃₇N₂O₈P644.66 645(M + H) 643(M − H) 667(M + Na) 757(M + TFA) 683(M + K) 108CH₂OPO(Me)(4-F—Ph) C₂₉H₃₂FN₂O₈P 586.55 587(M + H) 585(M − H) 609(M + Na)699(M + TFA) 109 CH₂OPO(n-hexyl)(4-F—Ph) C₃₄H₄₂FN₂O₈P 656.69 679(M + Na)655(M − H) 110 CH₂OPO(Me)(1-naphthyl) C₃₃H₃₅N₂O₈P 618.62 619(M + H)731(M + TFA) 641(M + Na) 111 CH₂O(6-Me-2-pyron-4-yl) C₂₈H₃₀N₂O₉ 538.55539(M + H) 112 CH₂O(4-coumarinyl) C₃₁H₃₀N₂O₉ 574.59 575(M + H) 537(M −H) 597(M + Na) 687(M + TFA) 113 CH₂O(2-Me-4-pyron-3-yl) C₂₈H₃₀N₂O₉538.55 539(M + H) 537(M − H) 561(M + Na) 651(M + TFA) 114CH₂O[1,2-diMe-4(1H)-pyridon-3-yl] C₂₉H₃₃N₃O₈ 551.59 552(M + H) 550(M −H) 115 CH₂O(3-flavonyl) C₃₇H₃₄N₂O₉ 650.68 651 (M + H) 649(M − H) 116CH₂O(4,6-diMe-2-pyrimidinyl) C₂₈H₃₂N₄O₇ 536.58 537(M + H) 535(M − H) 117CH₂O(4-CF₃-2-pyrimidinyl) C₂₇H₂₇F₃N₄O₇ 576.53 577(M + H) 575(M − H) 118CH₂S(4,6-diMe-2-pyrimidinyl) Chd 28H₃₂N₄O₆S 552.64 553(M + H) 551(M − H)575(M + Na) 665(M + TFA) 119 CH₂O(2,6-diMe-4-pyrimidinyl) C₂₈H₃₂N₄O₇536.58 537(M + H) 535(M − H) 120 CH₂O(6-CF₃-4-pyrimidinyl) C₂₇H₂₇F₃N₄O₇576.53 577(M + H) 575(M − H) 121 CH₂O(2-CF₃-4-pyrimidinyl) C₂₇H₂₇F₃N₄O₇576.53 577(M + H) 575(M − H) 122 CH₂S(2-imidazolyl) C₂₅H₂₈N₄O₆S 512.58513(M + H) 511(M − H) 625(M + TFA) 123 CH₂S(1-Me-2-imidazolyl)C₂₆H₃₀N₄O₆S 526.61 527(M + H) 525(M − H) 124 CH₂S(1H-1,2,4-triazol-3-yl)C₂₄H₂₇N₅O₆S 513.57 514(M + H) 512(M − H) 125CH₂S(4-Me-4H-1,2,4-triazol-3-yl) C₂₅H₂₉N₅O₆S 527.59 528(M + H) 526(M −H) 640(M + TFA) 126 CH₂S(1-Me-5-tetrazolyl) C₂₄H₂₈N₆O₆S 528.58 529(M +H) 527(M − H) 127 CH₂S(1-Ph-5-tetrazolyl) C₂₉H₃₀N₆O₆S 590.65 591(M + H)589(M − H) 128 CH₂S(5-Me-1,3,4-thiadiazol-2-yl) C₂₅H₂₈N₄O₆S₂ 544.64545(M + H) 543(M − H) 129 CH₂S(5-Ph-1,3,4-oxadiazol-2-yl) C₃₀H₃₀N₄O₇S590.65 591(M + H) 589(M − H) 613(M + Na) 703(M + TFA) 130CH₂S(3-Ph-1,2,4-oxadiazol-5-yl) C₃₀H₃₀N₄O₇S 590.65 591(M + H) 589(M − H)131 CH₂S(4-Ph-2-thiazolyl) C₃₁H₃₁N₃O₆S₂ 605.72 606(M + H) 604(M − H)628(M + Na) 132 CH₂S(4,5-diPh-2-imidazolyl) C₃₇H₃₆N₄O₆S 664.77 665(M +H) 663(M − H) 133 CH₂O(2-benzothiazolyl) C₂₉H₂₉N₃O₇S 563.62 564(M + H)562(M − H) 586(M + Na) 134 CH₂O(2-benzimidazolyl) C₂₉H₃₀N₄O₇ 546.58547(M + H) 545(M − H) 569(M + Na) 135 CH₂S(2-benzothiazolyl)C₂₉H₂₉N₃O₆S₂ 579.68 580(M + H) 578(M − H) 136 CH₂S(2-benzimidazolyl)C₂₉H₃₀N₄O₆S 562.64 563(M + H) 561(M − H) 675(M + TFA) 137CH₂O(2-quinolinyl) C₃₁H₃₁N₃O₇ 557.60 558(M + H) 556(M − H) 580(M + Na)670(M + TFA) 138 CH₂O(3-isoquinolinyl) C₃₁H₃₁N₃O₇ 557.60 558(M + H)556(M − H) 139 CH₂O(1-isoquinolinyl) C₃₁H₃₁N₃O₇ 557.60 558(M + H) 556(M− H) 580(M + Na) 670(M + TFA) 140 CH₂O(4-quinazolinyl) C₃₀H₃₀N₄O₇ 558.59559(M + H) 557(M − H) 141 CH₂O(8-quinolinyl) C₃₁H₃₁N₃O₇ 557.60 558(M +H) 556(M − H) 670(M + TFA) 142 CH₂O(3-Me-4-CO₂Et-isoxazol-5-yl)C₂₉H₃₃N₃O₁₀ 583.59 584(M + H) 582(M − H) 143CH₂O(1-Ph-3-CF₃-pyrazol-5-yl) C₃₂H₃₁F₃N₄O₇ 640.61 641(M + H) 639(M − H)144 CH₂O(5-CO₂Me-isoxazol-3-yl) C₂₇H₂₉N₃O₁₀ 555.54 556(M + H) 554(M − H)578(M + Na) 145 CH₂O(5-iPr-isoxazol-3-yl) C₂₈H₃₃N₃O₈ 539.58 540(M + H)538(M − H) 146 CH₂O(3-benzoisoxazolyl) C₂₉H₂₉N₃O₈ 547.56 548(M + H)546(M − H) 147 CH₂O(1-Me-5-CF₃-pyrazol-3-yl) C₂₇H₂₉F₃N₄O₇ 578.54 579(M +H) 577(M − H) 601(M + Na) 148 CH₂O(1-benzotriazolyl) C₂₈H₂₉H₅O₇ 547.57548(M + H) 660(M + TFA) 149 CH₂O(N-phthalimidyl) C₃₀H₂₉N₃O₉ 575.57576(M + H) 574(M + H) 688(M + TFA)

EXAMPLES 150–154

Starting from N-(valinyl)aspartic acid, β-tert-butyl, α-methyl ester(see Example 62, Part B), following the general methods described inExample 62, Parts C through G and utilizing (2-phenylphenoxy)acetic acidin place of (1-naphthyloxy)acetic acid in Part C, and the appropriateacid or phenol in place of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one inPart F, the compounds shown below in Table 6 were also prepared:

TABLE 6

MS(ES) Ex. B Formula MW pos. neg. 150 CH₂OCO(2,6-di-Cl—Ph) C₃₁H₃₀Cl₂N₂O₈629.49 629/631(M + H) 627/629(M − H) 651/653(M + Na) 741/743(M + TFA)667/669(M + K) 151 CH₂O(2,4,6-triF-Ph) C₃₀H₂₉F₃N₂O₇ 586.57 587(M + H)585(M − H) 609(M + Na) 699(M + TFA) 625(M + K) 152CH₂O(2,3,5,6-tetraF-Ph) C₃₀H₂₈F₄N₂O₇ 604.56 605(M + H) 603(M − H)717(M + TFA) 153 CH₂OPOPh₂ C₃₆H₃₇N₂O₈P 656.67 679(M + Na) 655(M − H)695(M + K) 769(M + TFA) 154 CH₂OPO(Me)Ph C₃₁H₃₅N₂O₈P 594.60 617(M + Na)593(M − H) 633(M + K) 707(M + TFA)

EXAMPLES 155–157

Starting from N-(valinyl)aspartic acid, β-tert-butyl, α-methyl ester(see Example 62, Part B), following the general methods described inExample 62, Parts C through G and utilizing (2-naphthyloxy)acetic acidin place of (1-naphthyloxy)acetic acid in Part C, and the appropriateacid or phenol in place of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one inPart F, the compounds shown below in Table 7 were also prepared:

TABLE 7

MS(ES) Ex. B Formula MW pos. neg. 155 CH₂OCO(2,6-di-Cl—Ph) C₂₉H₂₈Cl₂N₂O₈603.45 603/605(M + H) 601/603(M − H) 625/627(M + Na) 715/717(M + TFA)156 CH₂O(2,4,6-triF—Ph) C₂₈H₂₇F₃N₂O₇ 560.53 583(M + Na) 559(M − H)673(M + TFA) 157 CH₂O(2,3,5,6-tetraF—Ph) C₂₈H₂₆F₄N₂O₇ 578.52 601(M + Na)577(M − H) 891(M + TFA)

EXAMPLES 158–159

Starting from N-(valinyl)aspartic acid, β-tert-butyl, α-methyl ester(see Example 62, Part B), following the general methods described inExample 62, Parts C through G and utilizing 3-(1-naphthyloxy)propionicacid in place of (1-naphthyloxy)acetic acid in Part C, and theappropriate acid or phenol in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the compounds shownbelow in Table 8 were also prepared:

TABLE 8

MS(ES) Ex. B Formula MW pos. neg. 158 CH₂OCO(2,6-di-Cl—Ph) C₃₀H₃₀Cl₂N₂O₈617.48 617/619(M + H) 615/617(M − H) 639/641(M + Na) 729/731(M + TFA)159 CH₂O(1-Ph-5-CF₃- C₃₃H₃₃F₃N₄O₇ 654.64 677(M + Na) 653(M − H)pyrazol-3-yl 767(M + TFA)

EXAMPLE 160

(3S)-3-[N-(3′-(Phenoxy)Propionyl)Valinyl]Amino-5-(2,3,5,6-Tetrafluorophenoxy)-4-OxopentanoicAcid

Starting from N-(valinyl)aspartic acid, β-tert-butyl, α-methyl ester(see Example 62, Part B), following the general methods described inExample 62, Parts C through G and utilizing 3-(phenoxy)propionic acid inplace of (1-naphthyloxy)acetic acid in Part C, and2,3,5,6-tetrafluorophenol in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the title compound wasalso prepared. MS(ES) for C₂₅H₂₆F₄N₂O₇ (MW 542.48): positive 543(M+H),565(M+Na), 581(M+K); negative 541(M−H).

EXAMPLE 161

(3S)-3-[N-((2-Phenoxyphenyl)Acetyl)Leucinyl]Amino-5-(Diphenylphosphinyloxy)-4-OxopentanoicAcid

Part A: [(N-Benzyloxycarbonyl)Leucinyl]Aspartic Acid β-tert-Butyl,α-Methyl Ester

To a solution of (N-benzyloxycarbonyl)leucine, N-hydroxysuccinimideester (4.54 g, 12.5 mmol) and aspartic acid, β-tert-butyl, α-methylester hydrochloride (3.00 g, 12.5 mmol) in CH₂Cl₂ (20 mL) at roomtemperature under nitrogen was added N-methylmorpholine (1.65 mL, 15mmol). After stirring at room temperature for 18 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed with 5%KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to give the title compound (5.56 g, 99%)as viscous oil. TLC(EtOAc-hexane; 1:1) Rf=0.48.

Part B:(3S)-3-[N-((2-Phenoxyphenyl)Acetyl)Leucinyl]Amino-5-(Diphenylphosphinyloxy)-4-OxopentanoicAcid

Starting with [(N-benzyloxycarbonyl)leucinyl]aspartic acid,β-tert-butyl, α-methyl ester and following the methods described inExample 62, Parts B through G, utilizing (2-phenylphenoxy)acetic acid inplace of (1-naphthyloxy)acetic acid in Part C, and thediphenylphosphinic acid in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the title compound wasalso prepared. MS(ES) for C₃₇H₃₉N₂O₈P (MW 670.70): positive 671(M+H),693(M+Na); negative 669(M−H); 783(M+TFA).

EXAMPLES 162–164

Starting with [(N-benzyloxycarbonyl)leucinyl]aspartic acid,β-tert-butyl, α-methyl ester (see Example 161, Part A) and following themethods described in Example 62, Parts B through G, utilizing(2-phenylphenoxy)acetic acid in place of (1-naphthyloxy)acetic acid inPart C, and the appropriate acid or phenol in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the compounds shown inTable 9 were also prepared.

TABLE 9

MS(ES) Ex. B Formula MW pos. neg. 162 CH₂OCO(2,6-di-Cl—Ph) C₃₂H₃₂Cl₂N₂O₈643.52 665/667(M + Na) 641/643(M − H) 755/757(M + TFA) 163CH₂O(2,4,6-triF—Ph) C₃₁H₃₁F₃N₂O₇ 600.60 623(M + Na) 599(M − H) 713(M +TFA) 164 CH₂O(2,3,5,6-tetraF—Ph) C₃₁H₃₀F₄N₂O₇ 618.59 641(M + Na) 731(M +TFA)

EXAMPLE 165

(3S)-3-[N-((2′-Carboxy-1′-Naphthyloxy)Acetyl)Leucinyl]-Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid

Part A: (2-Carbo-tert-Butoxy-1-Naphthyloxy)Acetic Acid

To a suspension of 1-hydroxy-2-naphthoic acid (4.91 g, 26.1 mmol) intoluene (40 mL) at 80° C. (bath temp) under nitrogen was addeddimethylformamide di-tert-butyl acetal (25.0 mL, 104.3 mmol) dropwiseover 10 min. After stirring at 80° C. for an additional 30 min, thecooled mixture was diluted with Et₂O, washed successively with water,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and concentrated. The crude product was combined with that of asmaller run starting with 0.196 g of 1-hydroxy-2-naphthoic acid (total:5.106 g, 27 mmol) and purified by flash chromatography on silica geleluting with EtOAc-hexane (5:95) to give 2-carbo-tert-butoxy-1-naphthol(5.52 g, 83%) as a colorless oil. TLC(EtOAc-hexane; 1:9) Rf=0.68.

To a solution of 2-carbo-tert-butoxy-1-naphthol (4.00 g, 16.4 mmol) indimethylformamide (16 mL) at room temperature under nitrogen was addedmethyl bromoacetate (1.7 mL, 18 mmol) and potassium fluoride (2.85 g, 49mmol). After stirring at room temperature for 16 hrs, TLC showed thereaction was still incomplete. Potassium carbonate (3.0 g, 21.7 mmol)and additional methyl bromoacetate (1.5 mL, 15.8 mmol) were added andthe mixture heated to 60° C. (bath temp). After heating at 60° C. for 1hr, the mixture was partitioned between EtOAc-water. The organic phasewas washed with water (2×) and saturated NaCl solution, dried overanhydrous sodium sulfate and evaporated to an oil (6.17 g).TLC(EtOAc-hexane; 5:95) Rf=0.18.

The above crude product (6.17 g, ca 16.4 mmol) was taken up in dioxane(100 mL) and treated with 1.0 N LiOH solution (33 mL, 33 mmol). Afterstirring at room temperature for 1 hr, 100 mL of 1.0 N NaOH was addedand the mixture washed with Et₂O. The aqueous phase was acidified (pH 2)with conc HCl and extracted with EtOAc. The EtOAc extract was washedwith saturated NaCl solution, dried over anhydrous sodium sulfate andevaporated to give the title compound as a viscous oil (6.02 g). Thecrude product is used without further purification.

Part B:(3S)-3-[N-((2′-Carboxy-1′-Naphthyloxy)Acetyl)-Leucinyl]Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid

Starting with [(N-benzyloxycarbonyl)leucinyl]aspartic acid,b-tert-butyl, a-methyl ester (see Example 161, Part A) and following themethods described in Example 62, Parts B through G, utilizing(2-carbo-tert-butoxy-1-naphthyloxy)acetic acid in place of(1-naphthyloxy)acetic acid in Part C, and 2,6-dichlorobenzoic acid inplace of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the titlecompound was prepared. MS(ES) for C₃₁H₃₀Cl₂N₂O₁₀ (MW 661.49): positive661/663(M+H), 683/685(M+Na), 699/701(M+Na); negative 659/661(M−H).

EXAMPLES 166–167

Starting with [(N-benzyloxycarbonyl)leucinyl]aspartic acid,β-tert-butyl, α-methyl ester (see Example 161, Part A) and following themethods described in Example 62, Parts B through G, utilizing(2-carbo-tert-butoxy-1-naphthyloxy)acetic acid in place of(1-naphthyloxy)acetic acid in Part C, and the appropriate acid or phenolin place of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, thecompounds shown in Table 10 were also prepared.

TABLE 10

MS(ES) Ex. B Formula MW pos. neg. 166 CH₂OPOPh₂ C₃₆H₃₇N₂O₁₀P 688.67689(M + H) 687(M − H) 167 CH₂O(2,3,5,6-tetraF—Ph) C₃₀H₂₈F₄N₂Ohd 9 636.55637(M + H) 635(M − H) 659(M + Na) 675(M + K)

EXAMPLE 168

(3S)-3-[N-((2′-Carboxy-1′-Naphthyloxy)Acetyl)Valinyl]Amino-5-(2′-Fluorophenoxy)-4-OxopentanoicAcid

Part A: N-((2-Carbo-tert-Butoxy-1-Naphthyloxy)Acetyl)Valine Methyl Ester

To a solution of (2-carbo-tert-butoxy-1-naphthyloxy)acetic acid (1.20 g,3.97 mmol, see Example 165, Part A) and valine methyl esterhydrochloride (0.932 g, 5.56 mmol) in N-methylpyrrolidone(7.5mL)-CH₂Cl₂(7.5 mL) at room temperature under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(2.11 g, 5.56 mmol) and diisopropylethylamine (2.42 mL, 13.9 mmol).After stirring at room temperature for 3.5 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to dryness. The residue waspurified by flash chromatography on silica gel eluting with EtOAc-hexane(1:9 to 3:7) to give the title compound (1.40 g, 85%) as a colorlessoil. TLC(EtOAc-hexane; 1:1) Rf=0.76.

Part B:N-[N′-((2-Carbo-tert-Butoxy-1-Naphthyloxy)Acetyl)Valinyl]Aspartic acid,β-tert-Butyl, α-Methyl Ester

To a solution of N-((2-carbo-tert-butoxy-1-naphthyloxy)acetyl)valinemethyl ester (1.39 g, 3.34 mmol) in dioxane (15 mL) at room temperaturewas added 1.0 N LiOH solution (5.9 mL, 5.0 mmol). After stirring at roomtemperature for 2 hrs, the mixture was acidified (pH 2) with conc HCland extracted wit EtOAc. The EtOAc extract was washed with saturatedNaCl solution, dried over anhydrous sodium sulfate and evaporated togive the mono-carboxylic acid as a gummy solid (1.50 g). The crudeproduct is used without further purification.

To a solution of the above crude acid (1.50 g, ca 3.34 mmol) andaspartic acid, β-tert-butyl, α-methyl ester hydrochloride (0.800 g, 3.34mmol) in N-methylpyrrolidone(7.5 mL)-CH₂Cl₂(7.5 mL) at room temperatureunder nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(1.394 g, 3.67 mmol) and diisopropylethylamine (1.75 mL, 10 mmol). Afterstirring at room temperature for 16 hrs, the mixture was partitionedbetween EtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (1:9 to 1:1) togive the title compound (1.25 g, 64%) as a white foam.

Part C:(3S)-3-[N-((2′-Carboxy-1′-Naphthyloxy)Acetyl)-Valinyl]Amino-5-(2′-Fluorophenoxy)-4-OxopentanoicAcid

Starting withN-[N′-((2-carbo-tert-butoxy-1-naphthyloxy)acetyl)valinyl]aspartic acid,β-tert-butyl, α-methyl ester and following the methods described inExample 62, Parts D through G, utilizing 2-fluorophenol in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the title compound wasprepared. MS(ES) for C₂₉H₂₉FN₂O₉ (MW 568.55): positive 591(M+Na);negative 567(M−H).

EXAMPLES 169–171

Starting withN-[N′-((2-carbo-tert-butoxy-1-naphthyloxy)acetyl)valinyl]-aspartic acid,β-tert-butyl, α-methyl ester (see Example 168, Part B) and following themethods described in Example 62, Parts D through G, utilizing theappropiate acid or phenol in place of3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the compounds shown inTable 11 were also prepared.

TABLE 11

MS(ES) Ex. B Formula MW pos. neg. 169 CH₂O(2,3,5,6-tetraF—Ph)C₂₉H₂₆F₄N₂O₉ 622.53 645(M + Na) 621(M − H) 170 CH₂OCO(2,6-diCl—Ph)C₃₀H₂₈Cl₂N₂O₁₀ 647.46 669/671 (M + Na) 645/647 (M − H) 171 CH₂OPOPh₂C₃₅H₃₅N₂O₁₀P 674.64 697(M + Na) 673(M − H)

EXAMPLE 172

(3RS)-3-[N-((1′-Naphthyloxy)Acetyl)Cyclohexylalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid

Part A: (3S)-3-(N-Benzyloxycarbonyl)Amino-5-Bromo-4-Oxopentanoic Acidtert-Butyl Ester

A solution of (N-benzyloxycarbonyl)aspartic acid, β-tert-butyl ester(2.28 g, 7.06 mmol) and N-methylmorpholine (0.85 mL, 7.7 mmol) intetrahydrofuran (40 mL) at −10° C. (NaCl/ice bath) under nitrogen wastreated dropwise via syringe with isobutyl chloroformate (1.1 mL, 8.5mmol). After stirring at −10° C. for 20 min, the mixture was filtered(sinctered glass) into a pre-cooled receiver (ice bath) washing thefilter cake with additional tetrahydrofuran (approx. 10 mL). Thecombined filtrate was treated with excess diazomethane/Et₂O solution(prepared from 3.10 g, 21 mmol of 1-methyl-3-nitro-1-nitrosoguanidine,20 mL 40% KOH/10 ml Et₂O) at 0° C. (ice bath) under nitrogen. Afterstirring at 0° C. for 15 min and at room temperature for 30 min, thereaction mixture was again cooled to 0° C. and treated with 48% HBr(2.0mL, 12 mmol)/acetic acid(2.0 mL). After stirring at 0° C. for 15 min andat room temperature for 15 min, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, saturated NaHCO₃,and saturated NaCl solutions dried over anhydrous Na₂SO₄ and evaporatedto a dryness. Trituration with hexane gave the crude title compound(3.32 g) as a yellow oil. TLC(EtOAc-hexane; 1:1) Rf=0.60 (intermediatediazoketone Rf=0.52).

Part B:(3S,4RS)-3-(N-Benzyloxycarbonyl)Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

To a solution of(3S)-3-(N-benzyloxycarbonyl)amino-5-bromo-4-oxopentanoic acid tert-butylester (0.857 g, 2.14 mmol) and 2,3,5,6-tetrafluorophenol (0.410 g, 2.45mmol) in dimethylformamide (5.0 mL) at room temperature under nitrogenwas added potassium fluoride (0.40 g, 6.9 mmol). After stirring at roomtemperature for 16 hrs, the mixture was diluted with EtOAc, washed withsaturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to a to give the crude tetrafluorophenoxymethylketone (1.08 g, 98%) as a yellow, viscous oil. TLC(EtOAc-hexane; 1:1)Rf=0.57.

To a solution of the above crude ketone (1.08 g, ca 2.14 mmol) inethanol (10 mL) at 0° C. under nitrogen was added sodium borohydride(0.057 g, 1.5 mmol). After stirring at 0° C. for 1 hr. the excessreducing agent was discharged by treatment with acetone (1.0 mL), themixture concentrated and the residue partitioned between EtOAc-halfsaturated NH₄Cl solution. The organic phase was washed with saturatedNaHCO₃ and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to a dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (1:3) to give thetitle compound (1.012 g, 94%) as a colorless oil. TLC(EtOAc-hexane; 1:1)Rf=0.48.

Part C:(3S,4RS)-3-[(N-9-Fluorenylmethoxycarbonyl)Cyclohexylalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

To a solution of(3S,4RS)-3-(N-benzyloxycarbonyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (1.012 g, 2.08 mmol) in MeOH (25 mL) was added 10%Pd—C (0.30 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 4 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated to givethe crude amine (0.682 g, 93%) as a viscous oil. TLC(MeOH-CH₂Cl₂; 5:95)Rf=0.21.

To a solution of (N-9-fluorenylmethoxycarbonyl)cyclohexylalanine (0.763g, 1.94 mmol) in CH₂Cl₂(10 mL) at 0° C. (ice bath) under nitrogen wasadded hydroxybenzotriazole hydrate (0.282 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.447 g, 2.33 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with the above crude amine (0.682 g, ca 1.93 mmol) and thereacton allowed to warm to room temperature. After stirring at roomtemperature for 3 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was purified by flash chromatography eluting withEtOAc-hexane (1:2) to give the title compound (1.028 g, 73%) as yellowfoam. TLC(EtOAc-hexane; 1:2) Rf=0.20.

Part D:(3S,4RS)-3-[Cyclohexylalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

A mixture of(3S,4RS)-3-[(N-9-fluorenylmethoxycarbonyl)cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (1.028 g, 1.4 mmol) and 10%piperidine/dimethylformamide (3.0 mL) was stirred at room temperatureunder nitrogen for 2 hrs. The mixture was diluted with CH₂Cl₂, washedwith water and saturated NaHCO₃ solution, dried over anhydrous anhydrousNa₂SO₄ and evaporated to dryness. The residue was purified by flashchromatography eluting with isopropanol-CH₂Cl₂ (7:93) to give the titlecompound (0.561 g, 78%) as a white solid. TLC(MeOH-CH₂Cl₂; 5:95)Rf=0.43.

Part E:(3S,4RS)-3-[N-((1′-Naphthyloxy)Acetyl)Cyclohexylalaninyl]-Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

To a solution of (1-naphthyloxy)acetic acid (0.041 g, 0.20 mmol) and(3S,4RS)-3-[cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.092 g, 0.18 mmol) in CH₂Cl₂(5.0 mL) at 0° C.(ice bath) under nitrogen was added hydroxybenzotriazole hydrate (0.050g) followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (0.042 g, 0.22 mmol). After stirring at 0° C. for 10 minand at room temperature for 18 hrs, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to give the crude title compound (0.139 g) aswhite foam. TLC(EtOAc-hexane; 1:2) Rf=0.25.

Part F:(3RS)-3-[N-((1′-Naphthyloxy)Acetyl)Cyclohexylalaninyl]-Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoxypentanoicAcid tert-Butyl Ester

To a solution of crude(3S,4RS)-3-[N-((1′-naphthyloxy)-acetyl)-cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.139 g, ca 0.18 mmol) in CH₂Cl₂ (5 mL) at roomtemperature under nitrogen was added Dess-Martin Periodinane (0.099 g,0.23 mmol). After stirring at room temperature for 1.5 hrs, the mixturewas diluted with EtOAc, washed with 1.0 M Na₂S₂O₃, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa dryness. The residue was purified by flash chromatography on silicagel eluting with EtOAc-CH₂Cl₂-hexane (1:1:2) to give the title compound(0.086 g, 69% overall) as a colorless glass. TLC(EtOAc-hexane; 1:2) twospots (diastereomers) Rf=0.33 and 0.38. Note: racemization of the centeralpha to the ketone has apparently occurred at some point in thesynthesis.

Part G:(3RS)-3-[N-((1′-Naphthyloxy)Acetyl)Cyclohexylalaninyl]-Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoxypentanoicAcid

To a solution of(3RS)-3-[-((1′-naphthyloxy)-acetyl)-cyclohexylalaninyl]-amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoicacid, tert-butyl ester (0.086 g, 0.125 mmol) in CH₂Cl₂(2.0 mL) at roomtemperature under nitrogen was added trifluoroacetic acid (1.0 mL). Theresulting clear solution was stirred at room temperature for 1 hr,evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1) to give thetitle compound (0.066 g, 83%) as an off-white solid. MS(ES) forC₃₂H₃₂F₄N₂O₇ (MW 632.61): positive 633(M+H), 655(M+Na); negative631(M−H), 745(M+TFA).

EXAMPLES 173–175

Starting with(3S,4RS)-3-[cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (see Example 172, Part D) and following themethods described in Example 172, Parts E through G, the compounds shownbelow in Table 12 were also prepared:

TABLE 12

MS(ES) Ex. R¹ X n R² Formula MW pos. neg. 173 2-naphthyl O 0 HC₃₂H₃₂F₄N₂O₇ 632.61 633(M + H) 631(M − H) 655(M + Na) 745(M + TFA)671(M + K) 174 1-naphthyl O 1 H C₃₃H₃₄F₄N₂O₇ 646.63 647(M + H) 645(M −H) 669(M + Na) 759(M + TFA) 685(M + K) 175 (2-Ph)Ph O 0 H C₃₄H₃₄F₄N₂O₇658.65 659(M + H) 657(M − H) 681(M + Na) 771(M + TFA) 697(M + K)

EXAMPLE 176–177

Starting from (N-benzyloxycarbonyl)alanine and following the generalmethods described in Example 62, Parts A through G, utilizing either(2-phenylphenoxy)acetic acid or (2-naphthyloxy)acetic acid in place of(1-naphthyloxy)acetic acid in Part C, and 2,3,5,6-tetrafluorophenol inplace of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the compoundsshown below in Table 13 were also prepared.

TABLE 13

MS(ES) Ex. R¹ X n R² Formula MW pos. neg. 176 2-naphthyl O 0 HC₂₆H₂₂F₄N₂O₇ 550.46 551(M + H) 549(M − H) 573(M + Na) 663(M + TFA) 177(2-Ph)Ph O 0 H C₂₈H₂₄F₄N₂O₇ 576.50 577(M + H) 575(M − H) 599(M + Na)689(M + TFA)

EXAMPLE 178

(3S)-3-[N-α-((2′-Phenylphenoxy)Acetyl)Lysinyl]Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid Trifluoroacetate Salt

Starting from (N-α-benzyloxycarbonyl-N-ε-t-butoxycarbonyl)lysine andfollowing the general methods described in Example 62, Parts A throughG, utilizing (2-phenylphenoxy)acetic acid in place of(1-naphthyloxy)acetic acid in Part C, and 2,6-dichlorobenzoic acid inplace of 3-hydroxy-1,2,3-benzotriazin-4(3H)-one in Part F, the titlecompound was also prepared. MS(ES) for C₃₂H₃₃Cl₂N₃O₈ (MW 658.53):positive 658/660(M+H); negative 770/772(M+TFA).

EXAMPLE 179

(3S,2′RS,4′R)-3-[3′-((1-Naphthyloxy)Acetyl)-2′-Phenylthiazolidine-4′-Carbonyl]Amino-4-OxobutanoicAcid

Part A: (2RS,4R)-2-Phenylthiazolidine-4-Carboxylic Acid, Methyl Ester

To a suspension of L-cysteine methyl ester hydrochloride (1.717 g, 10mmol) in tetrahydrofuran (5.0 mL) at room temperature under nitrogen wasadded benzaldehyde (1.02 mL, 10 mmol) followed by triethylamine (4.2 mL,30 mmol). After stirring at room temperature for 3.5 hrs, the resultingmixture was filtered through a pad of silica gel eluting with EtOAc.Evaporation of the filtrate gave the title compound (1.95 g, 88%) as acolorless oil. TLC(EtOAc-hexane; 1:5) Rf=0.22.

Part B:(2RS,4R)-3-((1-Naphthyloxy)Acetyl)-2-Phenylthiazolidine-4-CarboxylicAcid, Methyl Ester

To a solution of (1-naphthyloxy)acetic acid (3.033 g, 15 mmol) andpyridine (1.46 mL, 18 mmol) in CH₂Cl₂ (50 mL) at room temperature undernitrogen was added cyanuric fluoride (1.52 mL, 18 mmol). After stirringat room temperature for 3 hrs, the mixture was filtered throughsinctered glass and the filtrate evaporated to a viscous oil. Theresidue was taken up in CH₂Cl₂ and diluted with CH₂Cl₂ to a total volumeof 15.0 mL (ca 1.0 mmol/ml).

To a solution of (2RS,4R)-2-phenylthiazolidine-4-carboxylic Acid, methylester (1.953 g, 8.7 mmol) and 2,6-di-tert-butylpyridine (1.95 mL, 8.7mmol) in CH₂Cl₂ (22 mL) at −30° C. (dry ice/acetonitrile bath) undernitrogen was added the above acid fluoride solution (9.0 mL, ca 9.0mmol). After stirring at −30° C. for 6 hrs, the mixture was allowed toslowly warm to room temperature. After stirring at room temperature for16 hrs, the mixture was concentrated and the residue partitioned betweenEtOAc-water. The EtOAc extract was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (1:3) to give thetitle compound (2.672 g, 75%) as a viscous oil.

Part C:(2RS,4R)-3-((1-Naphthyloxy)Acetyl)-2-Phenylthiazolidine-4-CarboxylicAcid

To a solution of(2RS,4R)-3-((1-naphthyloxy)acetyl)-2-phenylthiazolidine-4-carboxylicacid, methyl ester (2.50 g, 6.14 mmol) in dioxane(15 mL)-water(5.0 mL)at room temperature was added 1.0 N LiOH solution (6.75 mL, 6.75 mmol).After stirring at room temperature for 16 hrs, The mixture waspartitioned between EtOAc-5% KHSO₄. The organic phase was washed withsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated togive the title compound (2.42 g, 100%) as a viscous oil.TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.38.

Part D:(3S,2′RS,4′R)-3-[3′-((1-Naphthyloxy)Acetyl)-2′-Phenylthiazolidine-4′-Carbonyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of(2RS,4R)-3-((1-naphthyloxy)acetyl)-2-phenylthiazolidine-4-carboxylicacid (0.393 g, 1.00 mmol) in CH₂Cl₂(10 mL) at 0° C. (ice bath) undernitrogen was added hydroxybenzotriazole hydrate (0.161 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.288 g, 1.50 mmol). After stirring at 0° C. for 10 min,(3S)-amino-4-oxobutanoic acid tert-butyl ester semicarbazone,p-toluenesulfonate salt (0.402 g, 1.0 mmol) followed byN-methylmorpholine (0.12 mL, 1.0 mmol) was added. After stirring at 0°C. for 2 hrs and at room temperature for 18 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to dryness. The crude product waspurified by flash chromatography eluting with EtOAc to give the titlecompound (0.242 g, 40%) as a colorless foam. TLC(EtOAc) Rf=0.48.

Part E:(3S,2′RS,4′R)-3-[3′-((1-Naphthyloxy)Acetyl)-2′-Phenylthiazolidine-4′-Carbonyl]Amino-4-OxobutanoicAcid Semicarbazone

To a solution(3S,2′RS,4′R)-3-[3′-((1-naphthyloxy)acetyl)-2′-phenylthiazolidine-4′-carbonyl]amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.240 g, 0.40 mmol) in CH₂Cl₂ (2.6mL)-anisole(0.1 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (0.61 mL). The resulting solution was stirred atroom temperature for 18 hrs, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (0.195 g, 89%) as an off-white solid. TLC(MeOH-CH₂Cl₂;1:9) Rf=0.23.

Part F:(3S,2′RS,4′R)-3-[3′-((1-Naphthyloxy)Acetyl)-2′-Phenylthiazolidine-4′-Carbonyl]Amino-4-OxobutanoicAcid

A solution of(3S,2′RS,4′R)-3-[3′-((1-naphthyloxy)acetyl)-2′-phenylthiazolidine-4′-carbonyl]amino-4-oxobutanoicacid semicarbazone (0.195 g, 0.355 mmol) in 37% aqueousformaldehyde-acetic acid-methanol (1:1:3; v:v:v; 7.0 mL) was stirred atroom temperature under nitrogen for 18 hrs. The resulting solution wasconcentrated on a rotovap, diluted with water, frozen and lyophilized.The residue was taken up in MeOH, filtered through Celite and evaporatedto dryness. The residue was triturated with Et₂O to give the titlecompound (0.090 g, 51%) as a white solid. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.60.MS(ES) for C₂₆H₂₄N₂O₆S (MW 492.55): negative 491(M−H).

EXAMPLES 180–184

Following the general methods described in Example 179, Parts A throughF, utilizing the appropiate aldehyde in place of benzaldehyde in Part A,the compounds shown in Table 14 were also prepared. In the case ofExample 184, (4R)-thiazolidine-4-carboxylic acid, methyl ester wasprepared by treatment of (4R)-thiazolidine-4-carboxylic acid (Sigma)with HCl(g) in MeOH.

TABLE 14

MS(ES) Ex. R⁸ Formula MW pos. neg. 180 n-propyl C₂₃H₂₆N₂O₆S 458.53 —457(M − H) 181 n-hexyl C₂₆H₃₂N₂O₆S 500.61 501(M + H) 499(M − H) 539(M +Na) 182 iso- C₂₃H₂₆N₂O₆S 458.53 459(M + H) 457(M − H) propyl 183 cyclo-C₂₆H₃₀N₂O₆S 498.59 499(M + H) 497(M − H) hexyl 184 H C₂₀H₂₀N₂O₆S 416.45— 415(M − H)

EXAMPLE 185

(3S)-3-[N-((1-Naphthyloxy)Acetyl)-4′(trans)-Hydroxyprolinyl]Amino-4-OxobutanoicAcid

Part A: N-((1-Naphthyloxy)Acetyl)-4′(trans)-Hydroxytroline, Methyl Ester

To a solution of (1-naphthyloxy)acetic acid (1.87 g, 9.23 mmol) and4(trans)-hydroxyproline, methyl ester (1.34 g, 9.23 mmol) in CH₂Cl₂(92mL) at 0° C. (ice bath) under nitrogen was added hydroxybenzotriazolehydrate (1.48 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(2.65 g, 13.8 mmol). After stirring at 0° C. for 1 hr and at roomtemperature for 6 hrs, the mixture was concentrated and the residuepartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to give the title compound (2.59 g,85%) as a colorless oil. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.48.

Part B:(3S)-3-[N-((1-Naphthyloxy)Acetyl)-4′(trans)-Hydroxyprolinyl]Amino-4-OxobutanoicAcid

Starting with N-((1-naphthyloxy)acetyl)-4′(trans)-hydroxyproline, methylester and following the general methods described in Example 179, PartsC through F, the title compound was also prepared. MS(ES) for C₂₁H₂₂N₂O₇(MW 414.41): positive 415(M+H); negative 413(M−H).

EXAMPLE 186

(2′S,3S)-3-[N-((1-Naphthyloxy)Acetyl)Indoline-2′-Carbonyl]Amino-4-OxobutanoicAcid

Starting with (2S)-N-[(1-naphthyloxy)acetyl]indoline-2-carboxylic acid(see Example 61, Part B) and following the general methods described inExample 179, Parts D through F, the title compound was also prepared.TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.43. MS(ES) for C₂₅H₂₂N₂O₆ (MW446.46): positive 447(M+H); negative 445(M−H).

EXAMPLE 187

(3S)-3-[N-((3′-Trifluoromethylsulfonylamino-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Part A: (3-Trifluoromethylsulfonylamino-2-Naphthyloxy)Acetic Acidtert-Butyl Ester

To a solution of 3-amino-2-naphthol (0.796 g, 5.0 mmol) in acetone (25mL) at room temperature under nitrogen was added tert-butyl bromoacetate(0.89 mL, 5.0 mmol) and powdered anhydrous potassium carbonate (2.075 g,15 mmol). After stirring at room temperature for 18 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed with water(2×) and saturated NaCl solution, dried over anhydrous sodium sulfateand evaporated to an oil (1.37 g). TLC(EtOAc-hexane; 1:3) Rf=0.36 (Rf of3-amino-2-naphthol: 0.17).

To a solution of the crude product (1.37 g, ca 5.0 mmol) in CH₂Cl₂ (17mL) at −78° C. under nitrogen was added triethylamine (0.84 mL, 6.0mmol) followed by trifluoromethanesulfonic anhydride (1.00 mL, 6.0mmol). After stirring at −78° C. for 30 min, the mixture was allowed towarm to room temperature. After stirring at room temperature for 1 hr,the mixture was partitioned between EtOAc-water. The organic phase waswashed with 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions,dried over anhydrous Na₂SO₄ and evaporated to dryness. Trituration ofthe residue with hexane gave the title compound (1.46 g, 72%) as asolid. TLC(EtOAc-hexane; 1:3) Rf=0.42.

Part B: (3-Trifluoromethylsulfonylamino-2-Naphthyloxy)Acetic Acid

To a solution of (3-trifluoromethylsulfonylamino-2-naphthyloxy)aceticacid tert-butyl ester (1.46 g, 3.60 mmol) in CH₂Cl₂(37 mL)-anisole(0.1mL)-water(0.57 mL) at room temperature under nitrogen was addedtrifluroacetic acid (5.7 mL). After stirring at room temperature for 16hrs, the mixture was evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (1.17 g, 92%) as a solid. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.04.

Part C:(3S)-3-[N-((3′-Trifluoromethylsulfonylamino-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of (3-trifluoromethylsulfonylamino-2-naphthyloxy)aceticacid (0.175 g, 0.5 mmol) in N-methylpyrrolidone(1.0 mL)-CH₂Cl₂(5.0 mL)at 0° C. (ice bath) under nitrogen was added hydroxybenzotriazolehydrate (0.092 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.144 g, 0.75 mmol). After stirring for 15 min, the mixture was treatedwith (3S)-N-(valinyl)amino-4-oxobutanoic acid tert-butyl estersemicarbazone (0.165 g, 0.5 mmol, prepared by a method analogous to thatdescribed for N-(leucinyl)amino-4-oxobutanoic acid tert-butyl estersemicarbazone, see Example 1, Part B and Example 2, Part A) andN-methylmorpholine (0.066 mL, 0.6 mmol). After stirring at 0° C. for 2hrs and at room temperature for 16 hrs, the mixture was partitionedbetween EtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to dryness. Trituration of the residue withEt₂O-hexane gave the title compound (0.201 g, 61%) as a solid.TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.38.

Part D:(3S)-3-[N-((3′-Trfluoromethylsulfonylamino-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid Semicarbazone

A solution of(3S)-3-[N-((3′-trifluoromethylsulfonylamino-2′-naphthyloxy)acetyl)-valinyl]amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.201 g, 0.30 mmol) in 6.0NHCl/AcOH (3.0 mL) was stirred at room temperature under nitrogen for 1hr. The resulting mixture was evaporated to dryness and chased withtoluene. The residue was triturated with Et₂O to give the title compound(0.146 g, 80%) as a solid. TLC(MeOH-CH₂Cl₂; 1:9) Rf=0.08.

Part E:(3S)-3-[N-((3′-Trifluoromethylsulfonylamino-2′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

A solution of(3S)-3-[N-((3′-trifluoromethylsulfonyl-amino-2′-naphthyloxy)acetyl)valinyl]amino-4-oxobutanoicacid semicarbazone (0.146 g, 0.24 mmol) in MeOH-acetic acid-37% aqueousformaldhyde (3.0 mL, 3:1:1, v:v:v), was stirred at room temperatureunder nitrogen for 16 hrs. The mixture was concentrated, diluted withwater, frozen and lyophilized. The residue was taken up in methanol,filtered and evaporated to dryness. The residue was triturated with Et₂Oto give the title compound (0.103 g, 78%) as a solid. MS(ES) forC₂₂H₂₄F₃N₃O₈S (MW 547.50): negative 546(M−H).

EXAMPLE 188

(3S)-3-[N-((5′-Trifluoromethylsulfonylamino-1′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

Part A: (5-Trifluoromethylsulfonylamino-1-Naphthyloxy)Acetic Acid

To a solution of 5-amino-1-naphthol (0.790 g, 5.0 mmol) in acetone (25mL) at room temperature under nitrogen was added methyl bromoacetate(0.57 mL, 6.0 mmol) and powdered anhydrous potassium carbonate (2.075 g,15 mmol). After stirring at room temperature for 18 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed with water(2×) and saturated NaCl solution, dried over anhydrous sodium sulfateand evaporated to an oil (1.16 g).

To a solution of the above crude product (1.16 g, ca 5.0 mmol) in CH₂Cl₂(17 mL) at −78° C. under nitrogen was added triethylamine (0.84 mL, 6.0mmol) followed by trifluoromethanesulfonic anhydride (1.00 mL, 6.0mmol). After stirring at −78° C. for 30 min, the mixture was allowed towarm to room temperature. After stirring at room temperature for 1 hr,the mixture was partitioned between EtOAc-water. The organic phase waswashed with 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions,dried over anhydrous Na₂SO₄ and evaporated to dryness to give the crudesulfonamide (1.82 g, 100%) as a solid.

The above crude product was taken up in dioxane-water (16.7 mL, 3:1,v:v) and treated with 1.0N LiOH solution (11 mL, 11 mmol). Afterstirring at room temperature for 16 hrs, the mixture was acidified withconc HCl, and extracted with EtOAc. The EtOAc extract was washed withsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was triturated with Et₂O to give the title compound(1.27 g, 73%) as a solid.

Part B:(3S)-3-[N-((5′-Trifluoromethylsulfonylamino-1′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid tert-Butyl Ester Semicarbazone

To a solution of (5-trifluoromethylsulfonylamino-1-naphthyloxy)aceticacid (0.175 g, 0.5 mmol) and (3S)-N-(valinyl)amino-4-oxobutanoic acidtert-butyl ester semicarbazone (0.165 g, 0.5 mmol) inN-methylpyrrolidone(2.5 mL)-CH₂Cl₂ (2.5 mL) at 0° C. (ice bath) undernitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.165 g, 0.5 mmol) followed by diisopropylethylamine (0.17 mL, 0.98mmol). After stirring at 0° C. for 1 hr and at room temperature for 16hrs, the mixture was partitioned between EtOAc-water. The organic phasewas washed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness.Trituration of the residue with Et₂O-hexane gave the title compound(0.067 g, 20%) as a solid. TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.29.

Part C:(3S)-3-[N-((5′-Trfluoromethylsulfonylamino-1′-Naphthyloxy)Acetyl)Valinyl]Amino-4-OxobutanoicAcid

A solution of(3S)-3-[N-((5′-trifluoromethylsulfonylamino-1′-naphthyloxy)acetyl)-valinyl]amino-4-oxobutanoicacid tert-butyl ester semicarbazone (0.067 g, 0.10 mmol) in 6.0NHCl/AcOH (1.0 mL) was stirred at room temperature under nitrogen for 1hr. The resulting mixture was evaporated to dryness and chased withtoluene. TLC(AcOH-MeOH-CH₂Cl₂; 1:1:8) Rf=0.55.

A solution of the above crude product (ca 0.10 mmol) in MeOH-aceticacid-37% aqueous formaldhyde (3.3 mL, 3:1:1, v:v:v), was stirred at roomtemperature under nitrogen for 16 hrs. The mixture was concentrated,diluted with water, frozen and lyophilized. The residue was taken up inmethanol, filtered and evaporated to dryness. The residue was trituratedwith Et₂O to give the title compound (0.041 g, 75%) as a solid.TLC(AcOH-MeOH-CH₂Cl₂; 1:1:8) Rf=0.73. MS(ES) for C₂₂H₂₄F₃N₃O₈S (MW547.50): positive 570(M+Na); negative 546(M−H).

EXAMPLE 189

(3S)-3-[N-(4-(1′-Naphthyoxy)Butyryl)Valinyl]Amino-4-Oxobutanoic Acid

Part A: (3S)-3-[N-(9-Fluorenylmethoxycabonyl)Valinyl]Amino-4-OxobutanoicAcid (tert-Butyl) EsterSemicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetyl)]Aminomethylpolystrene

Aminomethylpolystryene resin (10.0 g, 100–200 mesh, 0.71 meq/g) wasplaced in a 200 mL filter tube equipped with a vacuum stopcock and glassfrit and washed successively with CH₂Cl₂(50 mL)/dimethylformamide(50mL), diisopropylethylamine(5 mL)/dimethylformamide(30 mL),dimethylformamide (2×50 mL) and tetrahydrofuran (30 mL). The resin wassuspended in tetrahydrofuran(20 mL)/N-methylpyrolidinone(20 mL) withnitrogen agitation through the bottom of the frit and treated withdiiospropylethylamine (1.9 mL, 10.9 mmol) and(3S)-3-(9-fluorenylmethoxycabonyl)amino-4-oxobutanoic acid(tert-butyl)ester semicarbazonyl-4-[2′-(4-ethyl-phenoxyacetic acid)](2.24 g, 3.56 mmol). After all of the solid had dissolved (approx. 10min), the mixture was treated with pyBOP[benzotriazolyloxy-tris(N-pyrolidinyl)phosphonium hexafluorophosphate,2.78 g, 5.34 mmol) in one portion. After mixing by nitrogen agitationfor 3 hrs, the supernatant was removed by suction and the resin washedsuccesively with tetrahydrofuran (2×50 mL), dimethylformamide (3×50 mL)and CH₂Cl₂ (2×50 mL). Unreacted amine groups were capped by treatmentwith a mixture of acetic anhydride(10 mL)/dimethylformamide(30mL)/diisopropylethylamine(1.0 mL). After mixing by nitrogen agitationfor 1 hr, the supernatant was removed by suction and the resin washedwith dimethylformamide(4×50 mL).

The resin was treated with piperidine(10 mL)/dimethylformamide(40 mL)and mixed by nitrogen agitation for 1 hr. The supernatant was removed bysuction and the resin washed with dimethylformamide(4×50 mL) andtetrahydrofuran (50 mL).

The resin was suspended in tetrahydrofuran(20mL)/N-methylpyrolidinone(20 mL), treated withN-(9-fluorenylmethoxycabonyl)valine (3.63 g, 10.7 mmol),diisopropylethylamine (5.7 mL, 32.7 mmol) and pyBOP (8.34 g. 16.0 mmol)and mixed by nitrogen agitation for 2.5 hrs. The supernatant was removedby suction and the resin washed succesively with dimethylformamide (3×40mL) and CH₂C₂ (3×40 mL), methanol (2×40 mL) and Et₂O (2×40 mL). Theresin was dried in vacuo to give the title product (12.69 g,quanitative). Based on the starting semicarbazone-acid, the resinloading was calculated as approximately 0.28 meq/g.

Part B: (3S)-3-[N-(4-(1′-Naphthyloxy)Butyryl)Valinyl]Amino-4-OxobutanoicAcid

An aliquot of the Part A resin (0.125 g, ca 0.035 mmol) was placed in a6 mL Supelco™ fitration tube equipped with a 20 μm polyethylene frit,treated with piperidine-dimethylformamide (1.0 mL, 1:4 v/v) and mixed onan orbital shaker for 1 hr. The supernatant was removed by suction andthe resin washed with dimethylformamide (4×1.0 mL) and CH₂Cl₂ (3×1.0mL). The resin was treated with 0.5M iPr₂NEt in N-methylpyrolidinone(0.40 mL, 0.20 mmol), 4-(1-naphthyloxy)butyric acid (0.0264 g, 0.115mmol) and 0.25M O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophate in N-methylpyrolidinone (0.40 mL, 0.10 mmol). Themixture was mixed on an orbital shaker under an nitrogen atmosphere for16 hrs. The supernatant was removed by suction and the resin washedsuccesively with dimethylformamide (3×1.0 mL) and CH₂Cl₂ (3×1.0 mL),methanol (2×1.0 mL) and Et₂O (2×1.0 mL).

The resin was treated with 1.0 mL of CH₂Cl₂ and allowed to re-swell for15 min. The solvent was removed by suction and the resin treated withtrifluoroacetic acid-CH₂Cl₂-anisole (1.0 mL, 4:3:1 v/v/v). After mixingon an orbital shaker under nitrogen for 5.5 hrs, the supernatant wasremoved by suction and the resin washed with CH₂Cl, (4×1.0 mL). Theresin was treated with 37% aqueous formaldehyde-aceticacid-tetrahydrofuran-trifluoroacetic acid (1.0 mL, 1:1:5:0.025 v/v/v/v)and mixed on an orbital shaker under nitrogen for 4.5 hrs. Thesupernatant was collected by suction, the resin washed withtetrahydrofuran (3×0.5 mL). The combined filtrates were blown down undernitrogen. The residue was taken up in methanol (0.5 mL), filtered andapplied directly to a 3 mL Supelco™ LC-18 reverse phase extraction tubewhich had been pre-conditioned with water, and eluted successively with3 mL each of 10% MeOH-water, 30% MeOH-water, 60% MeOH-water and 90%MeOH-water. The product-containing fractions (TLC) were combined andevaporated to dryness to give the title compound (0.0132 g, 88%) as acolorless glass. TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.22. MS(ES) forC₂₃H₂₈N₂O₆ (MW 428.48): positive 451 (M+Na), 467(M+K); negative427(M−H).

EXAMPLES 190–194

Starting with(3S)-3-[N-(9-fluorenylmethoxycabonyl)valinyl]amino-4-oxobutanoic acid(tert-butyl)estersemicarbazonyl-4-[2′-(4-ethyl-phenoxyacetyl)]aminomethylpolystrene (seeExample 189, Part A) and following the methods described in Example 189,Part B, the compounds shown below in Table 15 were also prepared:

TABLE 15

MS(ES) Ex. R¹ X n R² Formula MW pos. neg. 190 (2-t-Bu)Ph O 0 HC₂₁H₃₀N₂O₆ 406.48 429(M + Na) 405(M − H) 445(M + K) 191 (2-Ph)Ph O 0 HC₂₃H₂₆N₂O₆ 426.47 449(M + Na) 425(M − H) 465(M + K) 192 (2-Ph)Ph O 0 CH₃C₂₄H₂₈N₂O₆ 440.50 463(M + Na) 439(M − H) 193 (2-Ph)Ph O 1 H C₂₄H₂₈N₂O₆440.50 441(M + H) 439(M − H) 463(M + Na) 553(M + TFA) 479(M + K) 1941-naphthyl O 1 H C₂₂H₂₆N₂O₆ 414.46 415(M + H) 413(M − H) 437(M + Na)453(M + K)

EXAMPLE 195 Methyl4-(1H-1,2,3,4-tetrazole-5-yl)(2S)-2-[(phenylmethoxy)carbonylamino]butanoate

The title compound was prepared according to the literature (Tran ThachVan, et al, Tetrahedron, 1977, 33, 2299–2302).

EXAMPLE 196

To a stirred and cooled (−15 C) solution of methyl4-(1H-1,2,3,4-tetrazole-5-yl)(2S)-2-[(phenylmethoxy)carbonylamino]butanoate(4.0 g, 12.53 mmol) and triethylamine (2.62 ml, 18.79 mmol) in DCM (50ml) was added α-chloromethyl methyl ether (MOMCl, 1.43 ml, 18.79 mmol).The stirring was continued for 2.5 hours and by which time the cold bathwas warmed to 0 C. The solution was diluted with saturated aqueousNaHCO₃ (100 ml) and the layers were separated. The aqueous layer wasback extracted with DCM (100 ml). The combined organic layers were dried(Na₂SO₄) and evaporated. Flash chromatography of the residue over silicagel (4.5×18 cm), using 20, 40, 60, and 80% ethyl acetate in hexanes,gave compound 196-a (1.977 g, 43%) and compound 196-b (2.574 g, 56%).The structures of compounds were tentatively assigned based on protonNMR experiments according to literature (R. Raap, et al, Can. J. Chem.1968, 47, 813). Compound 196-a has: ¹H-NMR (CD₃OD, 300 MHz) δ 2.00–2.40(m, 2H), 3.00 (t, 2H), 3.40 (s, 3H), 3.70 (s, 3H), 4.20–4.35 (m, 1H),5.10 (s, 2H), 5.82 (s, 2H), 7.22–7.40 (m, 5H). Compound 196-b has:¹H-NMR (CD₃OD, 300 MHz) δ 2.08–2.50 (m, 2H), 3.05 (t, 2H), 3.32 (s, 3H),3.70 (s, 3H), 4.25–4.38 (m, 1H), 5.10 (s, 2H), 5.70 (s, 3H), 7.24–7.40(m, 5H).

EXAMPLE 197(2S)-4-[2-(methoxymethyl)(1,2,3,4-tetrazole-5-yl)]-2-[(phenylmethoxy)carbonylamino]butanoicacid

Aqueous LiOH (1.0 M, 7.88 ml) was added to a stirred solution ofcompound 196-a (1.91 g, 5.25 mmol) in dioxane (24 ml). After stirring atroom temperature for 1 hour, it was diluted with ethyl acetate andwashed with aqueous HCl (1.0 N, 20 ml). The organic layer was thenwashed with brine and dried (Na₂SO₄). Concentration under vacuo gave thetitle compound as an oil (1.98 g, quantitative). ¹H-NMR (CD₃OD, 300 MHz)δ 2.00–2.40 (m, 2H), 3.00 (t, 2H), 3.40 (s, 3H), 4.20–4.35 (m, 1H), 5.10(s, 2H), 5.82 (s, 2H), 7.20–7.40 (m, 5H).

EXAMPLE 198(2S)-4-[2-(methoxymethyl)(1,2,3,4-tetrazole-5-yl)]-2-[(phenylmethoxy)carbonylamino]butanoicacid

Aqueous LiOH (1.0 M, 10.3 ml) was added to a stirred solution of(2S)-4-[2-(methoxymethyl)(1,2,3,4-tetrazole-5-yl)]-2-[(phenylmethoxy)carbonylamino]butanoicacid (2.50 g, 6.88 mmol) in dioxane (30 ml). After stirring at roomtemperature for 1 hour, it was diluted with ethyl acetate and washedwith aqueous HCl (1.0 N, 20 ml). The organic layer was then washed withbrine and dried (Na₂SO₄). Concentration under vacuo gave the titlecompound as an oil (2.45 g, quantitative). ¹H-NMR (CD₃OD, 300 MHz) δ2.08–2.50 (m, 2H), 3.05 (t, 2H), 3.32 (s, 3H), 4.25–4.38 (m, 1H), 5.10(s, 2H), 5.70 (s, 3H), 7.24–7.40 (m, 5H).

EXAMPLE 199 N-[valinyl]aspartic acid, α-methyl, β-tert-butyl diester

HOBt (3.19 g, 20.8 mmol) and EDAC (5.60 g, 29.2 mmol) were added to astirred solution of N-carbobenzyloxycarbonyl valine (5.24 g, 20.8 mmol)in methylene chloride/DMF (60 ml/30 ml) at 0° C. under nitrogen. After15 min, aspartic acid α-methyl, β-tert-butyl diester (5.00 g, 20.8 mmol)was added as a solid followed neat 4-methylmorpholine (2.40 ml, 21.8mmol). After stirring at 0 C. for 1 hour and at room temperature for 5hours, the mixture was partitioned between ethyl acetate and 5% KHSO₄solution. The aqueous solution was back-extracted with ethyl acetate andthe combined extracts were washed with saturated NaHCO₃ and brine, driedover sodium sulfate, and concentrated to give a solid. Trituration withether afforded of N-[carbobenzyloxycarbonyl valinyl]aspartic acid,α-methyl, β-tert-butyl diester as a white solid (8.36 g, 92%). TLC(CH₂Cl₂/MeOH, 95/5): Rf=0.48.

A solution of the above product (4.00 g, 9.17 mmol) in 200 ml ofmethanol was stirred with palladium on activated carbon (0.45 g) underan atmosphere of hydrogen (1 atm) for 50 min. The reaction mixture wasthen filtered through a pad of Celite and the filter cake was washedwith methanol and methylene chloride. The filtrates were combined andconcentrated, and the residue was chased with methylene -chloride togive N-[valinyl]aspartic acid, α-methyl, β-tert-butyl diester as a whitesolid (2.75 g, 99%). TLC (CH₂Cl₂/MeOH, 95/5): Rf=0.10.

EXAMPLES 200–300

Utilizing the above intermediates, the compounds shown below in Table 16may also be prepared (in the following table, the “Via” columnindicates, when applicable, the starting compound from which the titlecompound is made):

TABLE 16 Exp. Compound Via Exp. Compound Via 200

— 201

13 202

201 203

202 204

203 205

204 206

205 207

204 208

207 209

204 210

209 211

204 212

211 213

195 214

213 215

214 216

215 217

216 218

217 219

218 220

219 221

220 222

215 223

222 224

223 225

224 226

198 227

226 228

227 229

228 230

229 231

228 232

231 233

232 234

233 235

234 236

231 237

236 238

237 239

227 240

239 241

240 242

241 243

242 244

243 245

244 246

243 247

246 248

243 249

248 250

243 251

250 252

239 253

252 254

253 255

254 256

253 257

256 258

253 259

258 260

253 261

260 262

198 263

262 264

263 265

264 266

265 267

263 268

267 269

268 270

263 271

270 272

271 273

197 274

273 275

274 276

275 277

276 278

274 279

278 280

279 281

280 282

281 283

280 284

283 285

280 286

285 287

280 288

287 289

278 290

289 291

290 292

291 293

292 294

293 295

292 296

295 297

292 298

297 299

292 300

299

Although the invention has been described with reference to the examplesprovided above, it should be understood that various modifications canbe made without departing from the spirit of the invention. Accordingly,the invention is limited only by the claims.

1. A method of treating arthritis, comprising administering to a patientin need thereof an effective amount of a pharmaceutical compositioncomprising a compound of the following formula:

wherein: n is 0 or 1; q is 1; X is O or NH; A is a natural or unnaturalamino acid of Formula IIa–i:

B is a hydrogen atom, a deuterium atom, C₁₋₁₀ straight chain or branchedalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, substitutednaphthyl, 2-benzoxazolyl, substituted 2-oxazolyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), (CH₂)_(m)heteroaryl, halomethyl, CO₂R¹³, CONR¹⁴R¹⁵,CH₂ZR¹⁶, CH₂OCO(aryl), CH₂OCO(substituted aryl), CH₂OCO(heteroaryl),CH₂OCO(substituted heteroaryl), or CH₂OPO(R¹⁷)R¹⁸, where Z is an oxygenor a sulfur atom, or B is a group of the Formula IIIa–c:

R¹ is substituted phenyl, naphthyl, or substituted naphthyl; R² ishydrogen, lower alkyl, (CH₂)_(m)CO₂R, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), or (CH₂)_(m)tetrazolyl; R³ is hydrogen orlower alkyl; and wherein: R⁴ is alkyl, cycloalkyl, phenyl, substitutedphenyl, (CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂,(CH₂)_(p)CO₂R³, (CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹²,(CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), or (CH₂)_(m)heteroaryl, wherein heteroarylincludes (but is not limited to) pyridyl, thienyl, furyl, thiazolyl,imidazolyl, pyrazolyl, isoxazolyl, pyrazinyl, pyrimidyl, triazinyl,tetrazolyl, and indolyl; R^(4a) is hydrogen, or methyl, or R⁴ and R^(4a)taken together are —(CH₂)_(d)— where d is an interger from 2 to 6; R⁵ isphenyl, substituted phenyl, (CH₂)_(p)phenyl, (CH₂)_(p)(substitutedphenyl), cycloalkyl, or benzofused cycloalkyl; R⁶ is hydrogen, alkyl,cycloalkyl, phenyl, substituted phenyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R⁷ is hydrogen, fluorine, oxo, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹, SR¹²,or NHCOR¹⁰; R⁸ is hydrogen, oxo, alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl); R⁹ isalkyl, cycloalkyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or COR¹⁰; R¹⁰is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), OR¹³, or NR¹⁴R¹⁵; R¹¹ is hydrogen, alkyl,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹² is alkyl, phenyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R¹³ is alkyl, cycloalkyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R¹⁴ is hydrogen, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, substituted naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹⁵ is hydrogen or alkyl; or R¹⁴ and R¹⁵ taken togetherform a five, six or seven membered carbocyclic or heterocyclic ring,such as morpholine or N-substituted piperazine; R¹⁶ is phenyl,substituted phenyl, naphthyl, substituted naphthyl, heteroaryl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or (CH₂)_(m)heteroaryl; R¹⁷ and R¹⁸ are independentlyalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, or phenylalkyl,substituted phenylalkyl, or (cycloalkyl)alkyl; R¹⁹ and R²⁰ areindependently hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(m)phenyl, or (CH₂)_(m)(substituted phenyl), or R¹⁹ and R²⁰ takentogether are —(CH═CH)₂—; R²¹ is hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl); R²², R²³ and R²⁴are independently hydrogen or alkyl; Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S; Y²is O or NR²⁴; Y³ is CH₂, O, or NR²⁴; a is 0 or 1 and b is 1 or 2,provided that when a is 1 then b is 1; c is 1 or 2, provided that when cis 1 then a is 0 and b is 1; m is 1, 2, 3 or 4; and p is 1 or 2; or apharmaceutically acceptable salt thereof, in combination with apharmaceutically acceptable carrier.
 2. A method of treating hepatitis,comprising administering to a patient in need thereof an effectiveamount of a pharmaceutical composition comprising a compound of thefollowing formula:

wherein: n is 0 or 1; q is 1; X is O or NH; A is a natural or unnaturalamino acid of Formula IIa–i:

B is a hydrogen atom, a deuterium atom, C₁₋₁₀ straight chain or branchedalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, substitutednaphthyl, 2-benzoxazolyl, substituted 2-oxazolyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substitUted phenyl), (CH₂)_(m)(1 or2-naphthyl), (CH₂)_(m)heteroaryl, halomethyl, CO₂R¹³, CONR¹⁴R¹⁵,CH₂ZR¹⁶, CH₂OCO(aryl), CH₂OCO(substituted aryl), CH₂OCO(heteroaryl),CH₂OCO(substituted heteroaryl), or CH₂OPO(R¹⁷)R¹⁸, where Z is an oxygenor a sulfur atom, or B is a group of the Formula IIIa–c:

R¹ is substituted phenyl, naphthyl, or substituted naphthyl; R² ishydrogen, lower alkyl, (CH₂)_(p)CO₂R³, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), or (CH₂)_(m)tetrazolyl; R³ is hydrogen orlower alkyl; and wherein: R⁴ is alkyl, cycloalkyl, phenyl, substitutedphenyl, (CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂,(CH₂)_(p)CO₂R³, (CH₂)_(p)OR¹¹, (CH)_(p)SR¹², (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or (CH₂)_(m)heteroaiyl, wherein heteroaryl includes (but isnot limited to) pyridyl, thienyl, furyl, thiazolyl, imidazolyl,pyrazolyl, isoxazolyl, pyrazinyl, pyrimidyl, triazinyl, tetrazolyl, andindolyl; R^(4a) is hydrogen, or methyl, or R⁴ and R^(4a) taken togetherare —(CH₂)_(d)— where d is an interger from 2 to 6; R⁵ is phenyl,substituted phenyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),cycloalkyl, or benzofused cycloalkyl; R⁶ is hydrogen, alkyl, cycloalkyl,phenyl, substituted phenyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl); R⁷ ishydrogen, fluorine, oxo, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹, SR¹², or NHCOR¹⁰; R⁸ ishydrogen, oxo, alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R⁹ is alkyl, cycloalkyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), or COR¹⁰; R¹⁰ is hydrogen, alkyl,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), OR, or NR¹⁴R¹⁵; R¹¹ is hydrogen, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl); R¹² isalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R¹³ is alkyl, cycloalkyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R¹⁴ is hydrogen, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, substituted naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹⁵ is hydrogen or alkyl; or R¹⁴ and R¹⁵ taken togetherform a five, six or seven membered carbocyclic or heterocyclic ring,such as morpholine or N-substituted piperazine; R¹⁶ is phenyl,substituted phenyl, naphthyl, substituted naphthyl, heteroaryl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or (CH₂)_(m)heteroaryl; R¹⁷ and R¹⁸ are independentlyalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, or phenylalkyl,substituted phenylalkyl, or (cycloalkyl)alkyl; R¹⁹ and R²⁰ areindependently hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(m)phenyl, or (CH₂)_(m)(substituted phenyl), or R¹⁹ and R²⁰ takentogether are —(CH═CH)₂—; R²¹ is hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl); R²², R²³ and R²⁴are independently hydrogen or alkyl; Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S; Y²is O or NR²⁴; Y³ is CH₂, O, or NR²⁴; a is 0 or 1 and b is 1 or 2,provided that when a is 1 then b is 1; c is 1 or 2, provided that when cis 1 then a is 0 and b is 1; m is 1, 2, 3 or 4; and p is 1 or 2; or apharmaceutically acceptable salt thereof, in combination with apharmaceutically acceptable carrier.